3 Department of Microbiology and Immunology, 6174 University Boulevard, University of British Columbia, Vancouver, British Columbia, V6T 1Z3 Canada, and 4 Program of Experimental Pathology, Aichi Cancer Center, Nagoya 464, Japan
Received on May 8, 2002; revised on June 18, 2002; accepted on June 23, 2002
![]() |
Abstract |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Key words: carbohydrate sulfation/CD44/glycosylation/inflammatory cytokines
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
CD44 is a widely expressed cell adhesion receptor that binds hyaluronan (HA), a nonsulfated GAG present in the extracellular matrix (ECM), and mediates cellcell and cell-ECM interactions (reviewed in Lesley et al., 1997; Lesley and Hyman, 1998
; Siegelman et al., 1999
; Johnson et al., 2000
). CD44 has been implicated in leukocyte adhesion to HEV (Jalkanen et al., 1986
, 1987) and extravasation at inflammatory sites (Camp et al., 1993
; DeGrendele et al., 1997
). Interestingly, CD44 isolated from peripheral blood leukocytes is sulfated (Jalkanen et al., 1988
). Although CD44 can exist as several isoforms due to alternative splicing, in leukocytes it is predominantly present as the standard isoform of ~85 kDa, referred to as CD44H, containing none of the alternatively spliced exons. CD44 can be modified by GAGs, including chondroitin sulfate (CS), heparan sulfate, and keratan sulfate (Takahashi et al., 1996
; Greenfield et al., 1999
). This typically results in the expression of higher-molecular-mass species of CD44 in the range of 150200 kDa (Jalkanen et al., 1988
). GAG-modified CD44 has been shown to bind components of the ECM, fibronectin (Jalkanen and Jalkanen, 1992
) and collagen (Carter and Wayner, 1988
; Ehnis et al., 1996
) as well as certain chemokines (Tanaka et al., 1993
) and growth factors (Bennett et al., 1995
), and can promote cell migration (Faassen et al., 1992
). CD44 is also modified with N- and O-linked glycans, and these oligosaccharides have been shown to play an important role in modulating the ability of CD44 to bind HA (Kincade et al., 1997
; Lesley et al., 1997
). However, the mechanisms regulating CD44 carbohydrate remodeling are still largely unknown.
A multitude of factors can affect the HA binding ability of CD44, such as cytoskeletal associations, oligomerization, isoform expression, and posttranslational modifications, such as N- and O-linked carbohydrate, GAG addition, sialylation, and sulfation (reviewed in Isacke, 1994; Kincade et al., 1997
; Lesley and Hyman, 1998
; Johnson et al., 2000
). However, the contribution and importance of these factors in the induction of HA binding by CD44 in response to physiological stimuli remain poorly defined. As with other leukocyte adhesion molecules, the ability of CD44 to bind its ligand, HA, is tightly regulated. Although CD44 is expressed in all leukocytes, most unstimulated cells do not bind HA. HA binding can be induced on cell activation by cytokines or antigen (Murakami et al., 1990
; Lesley et al., 1994
; Levesque and Haynes, 1997
). In monocytes, tumor necrosis factor
(TNF-
) can induce HA binding by CD44, and this has been correlated with changes in CD44 isoform expression, glycosylation and GAG addition, sialylation, and sulfation (Levesque and Haynes, 1996
, 1999; Maiti et al., 1998
; Katoh et al., 1999
; Jones et al., 2000
; Brown et al., 2001
). We have shown previously that TNF-
can induce CD44 sulfation in monocytes and in the SR91 myeloid cell line, and this correlated with the induction of HA binding and SR91 cell adhesion to an endothelial cell line (Maiti et al., 1998
; Brown et al., 2001
). Herein we identify the sulfated moiety of CD44 and determine the changes occurring to the sulfation of CD44 in response to the inflammatory cytokine TNF-
.
![]() |
Results |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
|
|
|
|
TNF- increases the presence of sulfated O-linked glycans on CD44
To directly assess the amount of sulfation due to O-linked glycosylation, the SR91 cells were treated with the O-linked glycosylation inhibitor, benzyl 2-acetamido-2-deoxy--D-galactopyranoside (BG), in the presence and absence of TNF-
. Initial results indicated no significant decrease in the overall sulfation of CD44, so the experiments were repeated in the presence of 2 mM xyloside to prevent GAG addition to the O-linked sites. Results are shown in Figure 6 and demonstrate that treatment with BG resulted in a slight decrease in apparent molecular mass of CD44. In the presence of xyloside, treatment with BG resulted in an additional decrease in the incorporation of [35S]sulfate into CD44 isolated from TNF-
-treated cells of 30 ± 19% (N = 4). Once again, the low level of incorporation of [35S]sulfate into CD44 from unstimulated SR91 cells made it difficult to assess the percentage change in sulfation, but from four experiments, no significant loss in sulfation was observed after BG treatment of unstimulated cells. This provides direct evidence for the presence of sulfated O-linked glycans on CD44 isolated from TNF-
-stimulated cells and provides data to support the notion that TNF-
enhances the sulfation of O-linked glycans, as well as the sulfation of N-linked glycans, on CD44 in SR91 cells. Considered together, the data indicate that CD44 is primarily sulfated on CS in unstimulated SR91 cells and that TNF-
increases the sulfation of both O- and N-linked glycans on CD44 and thus decreases the contribution of CS to the overall sulfation of CD44.
|
|
To determine if the 6-sulfo LacNAc/Lewis x determinant induced by TNF- was present on CD44, immunoprecipitates of cell surface CD44, isolated from unstimulated and TNF-
-stimulated SR91 cells that had been incubated in the presence or absence of 50 mM sodium chlorate were treated with neuraminidase, subjected to SDSPAGE, transferred to polyvinylidene difluoride (PVDF) membrane and subsequently probed with mAb AG107. Results showed that this epitope was not detected on CD44 from unstimulated SR91 cells but appeared on TNF-
stimulation and was abrogated by treatment of the cells with sodium chlorate (Figure 8). To determine if the 6-sulfo LacNAc/Lewis x epitope was present on N- or O-linked CD44 carbohydrate, western blots were performed with mAb AG107 after treatment of CD44 immunoprecipitates with PNGase F. Figure 8 demonstrates that PNGase F treatment of CD44 immunoprecipitates resulted in a substantial decrease in antibody reactivity (70 ± 10%, N = 3). This indicates that the 6-sulfo LacNAc/Lewis x determinant is present on CD44 and is expressed to a greater extent on N-linked glycans than on O-linked glycans.
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
We have been able to identify the different types and relative amounts of CD44 sulfation occurring on TNF- stimulation. However, there was a relatively large variability of [35S]sulfate incorporation into CD44 in response to TNF-
(2.5- ± 0.8-fold increase), as well as some variation in the percentage of sulfate due to GAGs and N- and O-linked glycans. This resulted in quite a range of percentages: 2040% for N-linked glycan sulfation, 525% for O-linked glycan sulfation, and 3458% for GAG sulfation. This variation may reflect a normal cellular variation in sulfation, glycosylation, and GAG synthesis and may also reflect variations in culture conditions. For example, the glucose levels present in the media have been reported to affect CD44 glycosylation (Zheng et al., 1997
) and ammonia, which can be released in the media from glutamine degradation, can down-regulate sialylation (Yang and Butler, 2000
).
We have previously shown that CD44 is sulfated in SR91 cells and in normal peripheral blood monocytes, and overall CD44 sulfation is increased on exposure to TNF- in both (Maiti et al., 1998
; Brown et al., 2001
). Here we show that CS and O- and N-linked glycans contribute to the overall sulfation of CD44 in SR91 cells and TNF-
can affect the amount and percentage of sulfation due to CS and N- and O-glycan sulfation in these cells. We have also found that CD44 is a major sulfated cell surface protein in another myeloid cell line, KG1a. Here, sulfation of CD44 was atttributed primarily to sulfation of N- and O-linked glycans (~30 and ~70% respectively, data not shown). In peripheral blood monocytes, preliminary data indicate the presence of CS and sulfated glycans on CD44 (Brown and Johnson, unpublished data). This indicates that the carbohydrate sulfation of CD44 is not restricted to SR91 cells but also occurs in myeloid cells. We had previously correlated the overall sulfation of CD44 with its ability to bind HA. However, here we have established that multiple sulfated moieties are present on CD44, indicating that the initial correlation was somewhat simplistic. It will now require further analysis to dissect the role and contribution of each sulfated moiety toward HA binding.
Further analysis using sulfated carbohydratespecific mAbs indicated that TNF--induced the expression of a 6-sulfo LacNAc or Lewis x determinant on the N-linked and (to a lesser extent) on the O-linked glycans of CD44. However, at present we do not know how much this type of sulfation contributes to the overall sulfation of CD44 glycans. In addition, because no antibodies were available that could detect 6-sulfo galactose or galactosamine, we cannot rule out the possibility that this type of sulfation could also be occurring. However, we have established the presence of 6-sulfo GlcNAc on both N- and O-linked CD44 glycans, and this raises the intriguing possibility that TNF-
may induce the expression or activation of a GlcNAc 6-O-sulfotransferase (GlcNAc6ST). However, TNF-
may also induce changes to CD44 glycosylation, which in turn may affect their sulfation, as changes in glycosylation have been reported to occur with CD44 upon cytokine treatment (Levesque and Haynes, 1999
; Cichy and Pure, 2000
).
There are now several glycosyl sulfotransferases identified, and most have very restricted substrate specificities (Hemmerich and Rosen, 2000). AG107 reactivity of isolated CD44 indicates that the 6-sulfo LacNAc/Lewis x epitope is present on N-linked glycans and, to a lesser extent, on O-linked glycans, implicating a GlcNAc6ST that can act on both N-linked and O-linked glycans. Putative candidates include the broadly expressed GlcNAc6ST-1 (Uchimura et al., 1998
) also known as CHST-2 or GST-2, and GlcNAc6ST-4 (Uchimura et al., 2000
) also known as C6ST-2 (Kitagawa et al., 2000
) or GST-5 (Bhakta et al., 2000
), which can transfer sulfate to both GlcNAc-Man and core 2 structures (Uchimura et al., 2000
), and can also sulfate 6-N-acetyl galactosamine to low levels (Kitagawa et al., 2000
). The nomenclature for the sulfotransferases is as described in Fukuda et al. (2001)
.
The MECA-79 mAb recognizes sulfated determinants present on HEV and has recently been shown to recognize the terminal epitope Galß1-4(sulfo-6)GlcNAcß1-3Galß1-3GalNAc present on core 1 but not core 2 mucin type O-glycans or N-glycans (Yeh et al., 2001). TNF-
-stimulated SR91 cells did not bind the MECA-79 mAb (data not shown), indicating that the sulfated determinant present on CD44 is distinct from that on HEV, which is recognized by MECA-79. AG107 reactivity to TNF-
-stimulated SR91 cells required the prior removal of sialic acid residues, implying that sialylation is occurring on the 6-sulfo LacNAc/Lewis x structures. However, the mAb G72, which recognizes
2
3 linked monosialylated 6-sulfo LacNAc/Lewis x, did not bind to TNF-
-stimulated SR91 cells. This suggests an alternative linkage for the terminal sialic acid or polysialylation, or alternatively, AG107 binding may be prevented by interference from neighboring sialic acid residues.
Here we have demonstrated the induction of a 6-sulfo LacNAc or Lewis x epitope on CD44 on SR91 cells in response to TNF-, and preliminary data suggest that the epitope is also induced on peripheral blood monocytes (Brown and Johnson, unpublished data). Sulfated sialyl Lewis x epitopes have been implicated in the initial adhesion or rolling of leukocytes on endothelial cells (Rosen, 1999
; Hemmerich and Rosen, 2000
), and sulfated CD44 has been implicated in regulating its adhesion to HA and to an endothelial cell line (Maiti et al., 1998
; Brown et al., 2001
). It is therefore possible that the induction of sulfated carbohydrates on CD44 augments leukocyte adhesion via either CD44 or a selectin ligand interaction. Now the sulfated moieties of CD44 have been identified, their role in leukocyte adhesion can be further explored.
SR91 cells express surface markers characteristic of myeloid cells and their progenitors. Early hematopoietic progenitor cells adhere to the bone marrow stroma, and CD44 has been implicated in this process (Legras et al., 1997). In particular, CS-modified CD44 and
4ß1 cooperate to mediate progenitor cell adhesion to fibronectin, an ECM component of the bone marrow stroma (Verfaillie et al., 1994
). Anti-CD44 antibodies can inhibit lympho- and myelopoiesis (Miyake et al., 1990
; Moll et al., 1998
), and CD44 knockout mice show anomalous myeloid progenitor distribution (Schmits et al., 1997
). It has also been shown that sulfated fucans can mobilize hematopoietic progenitor cells from the bone marrow (Sweeney et al., 2000
). Thus, it is possible that remodeling of CD44 by sulfated carbohydrates contributes to the regulation of myeloid cell adhesion during both development and inflammation.
![]() |
Materials and methods |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Antibodies and reagents
The rat mAb IM7 (IgG2b, Trowbridge et al., 1982), reactive against mouse and human CD44, was from J. Lesley and B. Hyman. The mouse mAb 3G12, directed against human CD44 (Dougherty et al., 1994
), was from G. Dougherty. The rat IgM mAb MECA-79 (Streeter et al., 1988
), was from S. Hemmerich. The following mouse IgM mAbs were also used: L4L4-8, anti-6,6'-disulfo LacNAc; AG107, anti-6-sulfo LacNAc/Lewis x; G72, anti-sialyl 6-sulfo LacNAc/Lewis x (Uchimura et al., 1998
); G270-16, anti-sialyl 6,6'-disulfo LacNAc/Lewis x; 2F3, anti-sialyl Lewis x, anti-sialyl 6-sulfo Lewis x, SU59, anti 3'-sulfo Lewis x (Ohmori et al., 1993
; Mitsuoka et al., 1998
; Izawa et al., 2000
). Fluoresceinated- or horseradish peroxidase (HRP)-conjugated anti-mouse IgG, anti-rat IgG, anti-mouse IgM, and streptavidin were from Jackson ImmunoResearch (West Grove, PA).
Cell surface biotinylation and neuraminidase treatment
Cell surface proteins were labeled using 0.5 mg NHS-LC-biotin (Pierce, Rockford, IL) for 3 x 107 cells in 1 ml phosphate buffered saline (PBS) 1% glucose for 30 min1 h at 4°C. Excess biotin was quenched with 1 mM lysine in PBS, and cells were washed three times in this buffer. Cells were lysed and processed for immunoprecipitations as described later. Alternatively, 107 cells/ml were incubated in the presence of 0.1 U/ml Vibrio cholerae neuraminidase (Roche Diagnostics, Laval, Quebec) in buffered medium (RPMI 1640/PBS, 1:1, pH 6.8) for 12 h at 37°C, washed, and then analyzed for antibody binding by flow cytometry or incubated at 4°C for 2 h with 1 ml CD44 mAb IM7 tissue culture supernatant prior to cell lysis and CD44 immunoprecipitation with Protein G Sepharose (Amersham Pharmacia Biotech, Baie dUrfé, Quebec) for 2 h at 4°C.
Biosynthetic labeling of SR91 with [35S]sodium sulfate or [35S]methionine-cysteine
Cells (1 x 106/ml) were incubated (2024 h) in sulfate-free RPMI medium (cell culture facility, University of California San Francisco) supplemented with 2 mM glutamine, 1 mM sodium pyruvate, and 10% fetal calf serum in the presence of 100 µCi/ml [35S]Na2SO4 (~1000 Ci/mmol, ICN Biomedicals, St Laurent, Quebec) or 40 µCi/ml [35S]methionine-cysteine protein labeling mix (1175 Ci/mmol; Easy Tag Express, NEN DuPont Canada, Markham, Ontario) in the culture medium containing 90% methionine-cysteine free RPMI 1640 medium (ICN) and 10% standard RPMI 1640 medium. In some cases, SR91 cells were also incubated for 2024 h with 10 ng/ml TNF- (R&D Systems, Minneapolis, MN) in the presence or absence of 50 mM sodium chlorate or 2 mM xyloside (Sigma) or both 2 mM xyloside and the O-glycosylation inhibitor, 2 mM BG (Sigma).
Immunoprecipitation
Labeled cells were rinsed three times in cold PBS before lysis in radio-immunoprecipitation assay lysis buffer containing the protease inhibitors (purchased from Sigma) phenylmethylsulfonyl fluoride (1 mM), leupeptin (1 µg/ml), aprotinin (1 µg/ml), and pepstatin (1 µg/ml). After centrifugation to remove particulate material, the resulting supernatants were precleared with Sepharose CL-4B beads (Sigma). The lysates were incubated with anti-CD44 mAb IM7 coupled to cyanogen bromideactivated Sepharose beads (Amersham Pharmacia; 4 mg/ml IM7-Sepharose, 50 µl/107 cells), for 2 h at 4°C. UltraLink-immobilized NeutravidinTM (Pierce) was used to precipitate biotinylated cell surface proteins. The beads were washed five times with lysis buffer, and immunoprecipitated material was eluted by boiling beads in sample buffer, resolved directly on 7.5% SDSPAGE, transferred to PVDF membrane (Millipore Canada, Mississauga, Ontario) and exposed to Kodak BioMax MR film (Interscience, Markham, Ontario) for approximately 4 days. Films were scanned and band intensities were determined with AlphaimagerTM software (Alpha Innotech, San Leandro, CA). Relative sulfation levels were expressed as the ratio of sulfated CD44 to immunoprecipitated CD44, which was determined by western blot analysis or in some cases by autoradiography of [35S]methionine-cysteine-labeled CD44.
Enzymatic treatment of immunoprecipitated CD44
CD44 from [35S]sulfate-labeled cells was immunoprecipitated, eluted from beads by boiling in sample buffer, precipitated with six volumes of acetone at 20°C, spun for 30 min at 16,000 x g, washed twice with acetone, and then resuspended in appropriate glycosidase buffer. CD44 immunoprecipitated from ~2 x 106 cells was incubated for 16 h at 37°C in 20 µl 50 mM sodium phosphate buffer, pH 7.5, containing 0.5% SDS, 1% NP40, and 1% ß-mercaptoethanol, with 500 U of PNGase F (New England Biolabs, Mississauga, Ontario) to remove N-linked glycans. Alternatively, CD44 immunoprecipitated from ~2 x 106 cells was incubated with 2 mU Proteus vulgaris chondroitin ABC lyase (Sigma) in 20 µl 50 mM TrisHCl, 50 mM sodium acetate, pH 8.0, for 16 h at 37°C to remove CS with 2 U/ml of keratanase from Pseudomonas sp. (Seikagaku America, Falmouth, MA) for 17 h at 37°C in 50 mM TrisHCl, pH 7.4, to remove keratan sulfate or with 0.4 U/ml of heparinase I or III from Flavobacterium heparinum (Sigma) in 25 mM sodium acetate, 100 mM NaCl, 5 mM calcium acetate, pH 7.0, for 16 h at 37°C to remove heparan sulfate. Samples were resolved on 7.5% SDSPAGE, transferred to PVDF membrane, and exposed to X-ray film.
ß-Elimination on PVDF membrane
[35S]sulfate-labeled CD44 was immunoprecipitated, separated by SDSPAGE, and transferred to PVDF membrane. CD44 associated GAGs and O-linked sugars were subjected to ß-elimination by incubating the PVDF membrane in 0.1 M NaOH for 16 h at 45°C (Duk et al., 1997). Membranes were rinsed twice in water, dried, and exposed to X-ray film.
Western blot
The dried PVDF membranes were incubated with 3G12, IM7, or AG107 hybridoma supernatants diluted 1:10 in 150 mM NaCl, 10 mM Tris, pH 7.5, 0.05% Tween 20 (TBST) containing 5% skim milk powder for 3G12 and IM7 and 3% bovine serum albumin for AG107. The blots were washed with TBST and incubated with the appropriate HRP-conjugated secondary antibody at 1:10,000 and washed again several times; proteins were detected by enhanced chemiluminescence (ECL; Amersham Pharmacia). In some cases, antibodies were stripped from membranes by incubation in 1% NP40, 0.1 M glycine, pH 3, for 1 h. Membranes were rinsed in TBST, dried, and stored until reprobed. When biotinylated cell surface antigens were used for immunoprecipitation, the blots were blocked for 1 h in 3% bovine serum albumin in TBST, washed with TBST, and incubated with 1:10,000 HRP-conjugated streptavidin (Jackson) for 30 min. Membranes were washed in TBST and the proteins detected by ECL.
Immunofluorescence staining and flow cytometry analysis
Cells were washed in ice-cold PBS containing 2% fetal calf serum and 0.1% NaN3 (FACS buffer), and 3 x 105 cells were incubated on ice for 30 min with 50 µl of 1:16 dilution of all hybridoma supernatants except IM7 and 3G12 supernatants, which were used undiluted, and the MECA-79 mAb, which was used at 10 µg/ml. The cells were washed with FACS buffer, incubated for 30 min with 1:100 fluoresceinated secondary antibody, and washed with FACS buffer. Labeled cells were resuspended in FACS buffer with propidium iodide (2 µg/ml), and flow cytometry analysis on gated live cells was performed on a FACScanTM (Becton Dickinson, Mississauga, Ontario) using CellQuestTM software.
![]() |
Acknowledgments |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
Abbreviations |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
Footnotes |
---|
2 To whom correspondence should be addressed; E-mail: pauline@interchange.ubc.ca
![]() |
References |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Bennett, K.L., Jackson, D.G., Simon, J.C., Tanczos, E., Peach, R., Modrell, B., Stamenkovic, I., Plowman, G., and Aruffo, A. (1995) CD44 isoforms containing exon v3 are responsible for the presentation of heparin-binding growth factor. J. Cell Biol., 128, 687698.[Abstract]
Bhakta, S., Bartes, A., Bowman, K.G., Kao, W.M., Polsky, I., Lee, J.K., Cook, B.N., Bruehl, R.E., Rosen, S.D., Bertozzi, C.R., and Hemmerich, S. (2000) Sulfation of N-acetylglucosamine by chondroitin 6-sulfotransferase 2 (GST-5). J. Biol. Chem., 275, 4022640234.
Bowman, K.G. and Bertozzi, C.R. (1999) Carbohydrate sulfotransferases: mediators of extracellular communication. Chem. Biol., 6, R9R22.[CrossRef][ISI][Medline]
Brockhausen, I. and Kuhn, W. (1997) Role and metabolism of glycoconjugate sulfation. Trends Glycosci. Glycotech., 9, 379398.[ISI]
Brown, K.L., Maiti, A., and Johnson, P. (2001) Role of sulfation in CD44-nediated hyaluronan binding induced by inflammatory mediators in human CD14+ peripheral blood monocytes. J. Immunol., 167, 53675374.
Bruehl, R.E., Bertozzi, C.R., and Rosen, S.D. (2000) Minimal sulfated carbohydrates for recognition by L-selectin and the MECA-79 antibody. J. Biol. Chem., 275, 3264232648.
Camp, R.L., Scheynius, A., Johansson, C., and Pure, E. (1993) CD44 is neccessary for optimal contact allergic responses but is not required for normal leukocyte extravasation. J. Exp. Med., 178, 497507.[Abstract]
Carter, W.G. and Wayner, E.A. (1988) Characterization of the class III collagen receptor, a phosphorylated, transmembrane glycoprotein expressed in nucleated human cells. J. Biol. Chem., 263, 41934201.
Cichy, J. and Pure, E. (2000) Oncostatin M and transforming growth factor-beta 1 induce post-translational modification and hyaluronan binding to CD44 in lung-derived epithelial tumor cells. J. Biol. Chem., 275, 1806118069.
DeGrendele, H.C., Estess, P., and Siegelman, M.H. (1997) Requirement for CD44 in activated T cell extravasation into an inflammatory site. Science, 278, 672675.
Dougherty, G.J., Cooper, D.L., Memory, J.F., and Chiu, R.K. (1994) Ligand binding specificity of alternatively spliced CD44 isoformsrecognition and binding of hyaluronan by CD44R1. J. Biol. Chem., 269, 90749078.
Duk, M., Ugorski, M., and Lisowska, E. (1997) Beta-elimination of O-glycans from glycoproteins transferred to immobilon P membranes: method and some applications. Anal. Biochem., 253, 98102.[CrossRef][ISI][Medline]
Ehnis, T., Dieterich, M., Bauer, M., Vonlampe, B., and Schuppan, D. (1996) A chondroitin/dermatan sulfate form of CD44 is a receptor for collagen xiv (undulin). Exp. Cell Res., 229, 388397.[CrossRef][ISI][Medline]
Faassen, A.E., Schrager, J.A., Klein, D.J., Oegema, T.R., Couchman, J.R., and McCarthy, J.B. (1992) A cell surface chondroitin sulfate proteoglycan, immunologically related to CD44, is involved in type I collagen-mediated melanoma cell motility and invasion. J. Cell Biol., 116, 521531.[Abstract]
Farzan, M., Mirzabekov, T., Kolchinsky, P., Wyatt, R., Cayabyab, M., Gerard, N.P., Gerard, C., Sodroski, J., and Hyeryun, C. (1999) Tyrosine sulfation of the amino terminus of CCR5 facilitates HIV-1 entry. Cell, 96, 667676.[ISI][Medline]
Farzan, M., Schnitzler, C.E., Vasilieva, N., Leung, D., Kuhn, J., Gerard, C., Gerard, N.P., and Choe, H. (2001) Sulfated tyrosines contribute to the formation of the C5a docking site of the human C5a anaphylatoxin receptor. J. Exp. Med., 193, 10591065.
Fukuda, M., Hiraoka, N., Akama, T.O., and Fukuda, M.N. (2001) Carbohydrate-modifying sulfotransferases: structure, function, and pathophysiology. J. Biol. Chem., 276, 4774747750.
Greenfield, B., Wang, W.C., Marquardt, H., Piepkorn, M., Wolff, E.A., Aruffo, A., and Bennett, K.L. (1999) Characterization of the heparan sulfate and chondroitin sulfate assembly sites in CD44. J. Biol. Chem., 274, 25112517.
Hemmerich, S. and Rosen, S.D. (2000) Carbohydrate sulfotransferases in lymphocyte homing. Glycobiology, 10, 849856.
Hemmerich, S., Butcher, E.C., and Rosen, S.D. (1994) Sulfation-dependent recognition of high endothelial venules (HEV)-ligands by L-selectin and MECA 79, and adhesion-blocking monoclonal antibody. J. Exp. Med., 180, 22192226.[Abstract]
Imai, Y., Lasky, L.A., and Rosen, S.D. (1993) Sulphation requirement for GlyCAM-1, an endothelial ligand for L-selectin. Nature, 361, 555557.[CrossRef][ISI][Medline]
Isacke, C.M. (1994) Commentary: the role of the cytoplasmic domain in regulating CD44 function. J. Cell Sci., 107, 23532359.
Izawa, M., Kumamoto, K., Mitsuoka, C., Kanamori, C., Kanamori, A., Ohmori, K., Ishida, H., Nakamura, S., Kurata-Miura, K., Sasaki, K., and others. (2000) Expression of sialyl 6-sulfo Lewis X is inversely correlated with conventional sialyl Lewis X expression in human colorectal cancer. Cancer Res., 60, 14101416.
Jalkanen, S. and Jalkanen, M. (1992) Lymphocyte CD44 binds the COOH-terminal heparin-binding domain of fibronectin. J. Cell Biol., 116, 817825.[Abstract]
Jalkanen, S., Jalkanen, M., Bargatze, R., Tammi, M., and Butcher, E.C. (1988) Biochemical properties of glycoproteins involved in lymphocyte recognition of high endothelial venules in man. J. Immunol., 141, 16151623.
Jalkanen, S.T., Bargatze, R.F., de los Toyos, J., and Butcher, E.C. (1987) Lymphocyte recognition of high endothelium: antibodies to distinct epitopes of an 8595 kD glycoprotein antigen differentially inhibit lymphocyte binding to lymph node, mucosal or synovial endothelial cells. J. Cell Biol., 105, 983990.[Abstract]
Jalkanen, S.T., Bargatze, R.F., Herron, L.R., and Butcher, E.C. (1986) A lymphoid cell surface glycoprotein involved in endothelial cell recognition and lymphocyte homing in man. Eur. J. Immunol., 16, 11951202.[ISI][Medline]
Johnson, P., Maiti, A., Brown, K.L., and Li, R. (2000) A role for the cell adhesion molecule CD44 and sulfation in leukocyte-endothelial cell adhesion during an inflammatory response? Biochem. Pharm., 59, 455465.[CrossRef][ISI][Medline]
Jones, M., Tussey, L., Athanasou, N., and Jackson, D.G. (2000) Heparan sulfate proteoglycan isoforms of the CD44 hyaluronan receptor induced in human inflammatory macrophages can function as paracrine regulators of fibroblast growth factor action. J. Biol. Chem., 275, 79647974.
Katoh, S., Miyagi, T., Taniguchi, H., Matsubara, Y., Kadota, J., Tominaga, A., Kincade, P.W., Matsukura, S., and Kohno, S. (1999) Cutting edge: an inducible sialidase regulates the hyaluronic acid binding ability of CD44-bearing human monocytes. J. Immunol., 162, 50585061.
Kincade, P.W., Zheng, Z., Katoh, S., and Hanson, L. (1997) The importance of cellular environment to function of the CD44 matrix receptor. Curr. Opin. Cell Biol., 9, 635642.[CrossRef][ISI][Medline]
Kitagawa, H., Fujita, M., Ito, N. , and Sugahara, K. (2000) Molecular cloning and expression of a novel chondroitin 6-O-sulfotransferase. J. Biol. Chem., 275, 2107521080.
Kjellen, L. and Lindahl, U. (1991) Proteoglycans: structures and interactions. Annu. Rev. Biochem., 60, 443475.[CrossRef][ISI][Medline]
Klingemann, H.G., Gong, H.J., Maki, G., Horsman, D.E., Dalal, B.I., and Phillips, G.L. (1994) Establishment and characterization of a human leukemic cell line (SR-91) with features suggestive of early hematopoietic progenitor cell origin. Leuk. Lymph., 12, 463470.[ISI][Medline]
Legras, S., Levesque, J.P., Charrad, R., Morimoto, K., Le Bousse, C., Clay, D., Jasmin, C., and Smadja-Joffe, F. (1997) CD44-mediated adhesiveness of human hematopoietic progenitors to hyaluronan is modulated by cytokines. Blood, 89, 19051914.
Lesley, J. and Hyman, R. (1998) CD44 structure and function. Front. Biosci., 3, 616630.
Lesley, J., Howes, N., Perschl, A., and Hyman, R. (1994) Hyaluronan binding function of CD44 is transiently activated on T cells during an in vivo immune response. J. Exp. Med., 180, 383387.[Abstract]
Lesley, J., Hyman, R., English, N., Catterall, J.B., and Turner, G.A. (1997) CD44 in inflammation and metastasis. Glycoconj. J., 14, 611622.[CrossRef][ISI][Medline]
Levesque, M.C. and Haynes, B.F. (1996) In vitro culture of human peripheral blood monocytes induces hyaluronan binding and up-regulates monocyte variant CD44 isoform expression. J. Immunol., 156, 15571565.[Abstract]
Levesque, M.C. and Haynes, B.F. (1997) Cytokine induction of the ability of human monocyte CD44 to bind hyaluronan is mediated primarily by TNF-alpha and is inhibited by IL-4 and IL-13. J. Immunol., 159, 61846194.[Abstract]
Levesque, M.C. and Haynes, B.F. (1999) TNF alpha and IL-4 regulation of hyaluronan binding to monocyte CD44 involves posttranslational modification of CD44. Cell. Immunol., 193, 209218.[CrossRef][ISI][Medline]
Lin, X. and Perrimon, N. (1999) Dally cooperates with Drosophila frizzled 2 to transduce wingless signalling. Nature, 400, 281284.[CrossRef][ISI][Medline]
Lin, X., Buff, E.M., Perrimon, N., and Michelson, A.M. (1999) Heparan sulfate proteoglycans are essential for FGF receptor signaling during Drosophila embryonic development. Development, 126, 37153723.
Maiti, A., Maki, G., and Johnson, P. (1998) TNF- induction of CD44-mediated leukocyte adhesion by sulfation. Science, 282, 941943.
Mitsuoka, C., Sawada-Kasugai, M., Ando-Furui, K., Izawa, M., Nakanishi, H., Nakamura, S., Ishida, H., Kiso, M., and Kannagi, R. (1998) Identification of a major carbohydrate capping group of the L-selectin ligand on high endothelial venules in human lymph nodes as 6-sulfo sialyl Lewis X. J. Biol. Chem., 273, 1122511233.
Miyake, K., Medina, K.L., Hayashi, S.-I., Ono, S., Hamaoka, T., and Kincade, P.W. (1990) Monoclonal antibodies to Pgp-1/CD44 block lympho-hemopoiesis in long-term bone marrow cultures. J. Exp. Med., 171, 477488.[Abstract]
Moll, J., Khaldoyanidi, S., Sleeman, J.P., Achtnich, M., Preuss, I., Ponta, H., and Herrlich, P. (1998) Two different functions for CD44 proteins in human myelopoiesis. J. Clin. Invest., 102, 10241034.
Murakami, S., Miyake, K., June, C.H., Kincade, P.W., and Hodes, R.J. (1990) IL-5 induces a Pgp-1 (CD44) bright B cell subpopulation that is highly enriched in proliferative and Ig secretory activity and binds to hyaluronate. J. Immunol., 145, 36183627.
Ohmori, K., Takada, A., Ohwaki, I., Takahashi, N., Furukawa, Y., Maeda, M., Kiso, M., Hasegawa, A., Kannagi, M., and Kannagi, R. (1993) A distinct type of sialyl Lewis X antigen defined by a novel monoclonal antibody is selectively expressed on helper memory T cells. Blood, 82, 27972805.[Abstract]
Pouyani, T. and Seed, B. (1995) PSGL-1 recognition of P-selectin is controlled by a tyrosine sulfation consensus at the PSGL-1 amino terminus. Cell, 83, 333343.[ISI][Medline]
Rosen, S.D. (1999) Endothelial ligands for L-selectinfrom lymphocyte recirculation to allograft rejection. Am. J. Pathol., 155, 10131020.
Sako, D., Comess, K.M., Barone, K.M., Camphausen, R.T., Cumming, D.A., and Shaw, G.D. (1995) A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding. Cell, 83, 323331.[ISI][Medline]
Sassetti, C., Tangemann, K., Singer, M.S., Kershaw, D.B., and Rosen, S.D. (1998) Identification of podocalyxin-like protein as a high endothelial venule ligand for L-selectinparallels to CD34. J. Exp. Med., 187, 19651975.
Schmits, R., Filmus, J., Gerwin, N., Senaldi, G., Kiefer, F., Kundig, T., Wakeham, A., Shahinian, A., Catzavelos, C., Rak, J., and others. (1997) CD44 regulates hematopoietic progenitor distribution, granuloma formation and tumorigenicity. Blood, 90, 22172233.
Sen, J., Goltz, J.S., Stevens, L., and Stein, D. (1998) Spatially restricted expression of pipe in the Drosophila egg chamber defines embryonic dorsal-ventral polarity. Cell, 95, 471481.[ISI][Medline]
Siegelman, M.H., DeGrendele, H.C., and Estess, P. (1999) Activation and interaction of CD44 and hyaluronan in immunological systems. J. Leukoc. Biol., 66, 315321.[Abstract]
Streeter, P.R., Rouse, B.T., and Butcher, E.C. (1988) Immunohistologic and functional characterization of a vascular addressin involved in lymphocyte homing into peripheral lymph nodes. J. Cell Biol., 107, 18531862.[Abstract]
Sweeney, E.A., Priestley, G.V., Nakamoto, B., Collins, R.G., Beaudet, A.L., and Papayannopoulou, T. (2000) Mobilization of stem/progenitor cells by sulfated polysaccharides does not require selectin presence. Proc. Natl Acad. Sci. USA, 97, 65446549.
Takahashi, K., Stamenkovic, I., Cutler, M., Dasgupta, A., and Tanabe, K.K. (1996) Keratan sulfate modification of CD44 modulates adhesion to hyaluronate. J. Biol. Chem., 271, 94909496.
Tanaka, Y., Adams, D.H., Hubscher, S., Hirano, H., Siebenlist, U., and Shaw, S. (1993) T-cell adhesion induced by proteoglycan-immobilized cytokine MIP-1beta. Nature, 361, 7982.[CrossRef][ISI][Medline]
Trowbridge, I.S., Lesley, J., Schulte, R., Hyman, R., and Trotter, J. (1982) Biochemical characterization and cellular distribution of a polymorphic, murine cell-surface glycoprotein expressed on lymphoid tissues. Immunogenetics, 15, 299312.[ISI][Medline]
Uchimura, K., Fasakhany, F., Kadomatsu, K., Matsukawa, T., Yamakawa, T., Kurosawa, N., and Muramatsu, T. (2000) Diversity of N-acetylglucosamine-6-O-sulfotransferases: molecular cloning of a novel enzyme with different distribution and specificities. Biochem. Biophys. Res. Commun., 274, 291296.[CrossRef][ISI][Medline]
Uchimura, K., Muramatsu, H., Kadomatsu, K., Fan, Q.W., Kurosawa, N., Mitsuoka, C., Kannagi, R., Habuchi, O., and Muramatsu, T. (1998) Molecular cloning and characterization of an N-acetylglucosamine-6-O-sulfotransferase. J. Biol. Chem., 273, 2257722583.
Verfaillie, C.M., Benis, A., Iida, J., Mcglave, P.B., and McCarthy, J.B. (1994) Adhesion of committed human hematopoietic progenitors to synthetic peptides from the C-terminal heparin-binding domain of fibronectin: cooperation between the integrin alpha 4 beta 1 and the CD44 adhesion receptor. Blood, 84, 18021811.
Wilkins, P.P., Moore, K.L., McEver, R.P., and Cummings, R.D. (1995) Tyrosine sulfation of P-selectin glycoprotein ligand-1 is required for high affinity binding to P-selectin. J. Biol. Chem., 270, 2267722680.
Yang, M. and Butler, M. (2000) Effect of ammonia on the glycosylation of human recombinant erythropoietin in culture. Biotechnol. Prog., 16, 751759.[CrossRef][ISI][Medline]
Yeh, J.C., Hiraoka, N., Petryniak, B., Nakayama, J., Ellies, L.G., Rabuka, D., Hindsgaul, O., Marth, J.D., Lowe, J.B., and Fukuda, M. (2001) Novel sulfated lymphocyte homing receptors and their control by a Core 1 extension beta 1, 3-N-acetylglucosaminyltransferase. Cell, 105, 957969.[CrossRef][ISI][Medline]
Zheng, Z., Cummings, R.D., Pummill, P.E., and Kincade, P.W. (1997) Growth as a solid tumor or reduced glucose concentrations in culture reversibly induce CD44-mediated hyaluronan recognition by chinese hamster ovary cells. J. Clin. Invest., 100, 12171229.