From the Department of Biochemistry and Molecular
Biology and the Oklahoma Center for Medical Glycobiology, University of
Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190 and the
§ Department of Pathology, University of Rochester School of
Medicine, Rochester, New York 14642
Received for publication, November 11, 2002, and in revised form, December 19, 2002
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Hyaluronan (HA) and chondroitin sulfate clearance
from lymph and blood is mediated by the hyaluronan receptor for
endocytosis (HARE). The purification and molecular cloning (Zhou, B.,
Weigel, J. A., Saxena, A., and Weigel, P. H. (2002)
Mol. Biol. Cell 13, 2853-2868) of this cell surface
receptor were finally achieved after we developed monoclonal antibodies
(mAbs) against HARE. There are actually two independent isoreceptors
for HA, which in rat are designated the 175-kDa HARE and 300-kDa HARE.
Only one mAb (number 174) effectively and completely blocked the
specific uptake of 125I-HA at 37 °C by rat liver
sinusoidal endothelial cells. 125I-HA binding to both the
175-kDa and 300-kDa HARE proteins in a ligand blot assay was almost
completely inhibited by <1 µg/ml mAb-174, whereas mouse IgG had
little or no effect. MAb-174 also performed very well in Western
analysis, indirect fluorescence microscopy, and a variety of
immuno-procedures. Immunohistochemistry using mAb-174 localized HARE to
the sinusoidal cells of rat liver, spleen, and lymph node. Western
analysis using mAb-174 revealed that the sizes of both HARE
glycoproteins were the same in these three tissues. 125I-HA
was taken up and degraded by excised rat livers that were continuously
perfused ex vivo with a recirculating medium. This HA
clearance and metabolism by liver, which is a physiological function of
HARE, was very effectively blocked by mAb-174 but not by mouse IgG. The
results indicate that mAb-174 will be a useful tool to study the
functions of HARE and the physiological significance of HA clearance.
After Meyer and Palmer (1) discovered hyaluronan
(HA),1 it was found to be a
component of essentially all vertebrate extracellular matrices (ECMs).
Fibroblasts, keratinocytes, chondrocytes, and other cells continuously
synthesize and secrete HA, which is a linear polymer with a native
molecular mass that may exceed 107 Da and is
composed of the repeating disaccharide
2-deoxy,2-acetamido-D-glucopyranosyl- Cell surface HA receptors identified to date include CD44, RHAMM
(CD168), ICAM-1 (CD54), LYVE-1 (5), and an endocytic receptor that is
specific for HA and chondroitin sulfate. This latter HA receptor is
expressed in LECs, which remove HA and chondroitin sulfate from the
blood (2, 17-20). Because this endocytic receptor is also in other
tissues and is not a liver-specific HA receptor, it was renamed HARE,
the HA receptor for endocytosis
(21). Unlike the other cell surface receptors for HA, HARE mediates the
rapid endocytosis of HA and chondroitin sulfate via the clathrin-coated
pit pathway (19, 22). HARE is, therefore, similar in its mode of action
to the transferrin, asialoglycoprotein, mannose, and low density
lipoprotein receptors (23).
After the discovery that liver is responsible for HA clearance from the
blood (reviewed in Ref. 2), Deaciuc et al. (24, 25)
demonstrated the ability of isolated perfused rat liver to take up HA.
This uptake process was saturable at an HA concentration of ~0.15
µg/ml with a steady-state uptake rate of ~10 µg HA/h/g wet weight
of liver. Deaciuc et al. (24, 25) also showed that galactosamine-induced hepatitis in rats was associated with elevated plasma HA levels and decreased HA clearance ability by perfused liver.
These and similar observations by other investigators led to the
realization that the HA clearance ability of liver can indicate the
functional status of LECs and, thus, the general health of the liver.
For example, Itasaka et al. (26) and Rao et al.
(27) found that the HA clearance function was a useful parameter for
predicting the likely success of human liver transplantation.
We previously identified two large membrane proteins of ~175 kDa and
~300 kDa in rat LECs that were specifically labeled with a
photoaffinity derivative of HA (28) and retained specific HA binding
activity in a novel ligand blot assay following SDS-PAGE and
renaturation (29). The development of specific mAbs raised against the
175-kDa protein enabled these two HARE proteins to be purified from
isolated rat LECs (30) and more recently from human spleen (31). After
cloning the cDNA for the rat 175HARE, we stably expressed the
recombinant protein in SK-Hep-1 cells and found that this smaller HARE
species can function as an endocytic recycling receptor with
specificity for HA and chondroitin sulfate. The native rat 175HARE is
derived by proteolysis from a larger precursor protein (32), probably
the largest subunit of the 300HARE. The evidence so far indicates that
the 175- and 300-kDa proteins are closely related but functionally
independent HARE species.
We noted previously that one of the mAbs generated to the rat HARE,
designated mAb-174, inhibits 125I-HA endocytosis by
isolated rat LECs and inhibits 125I-HA binding to both HARE
species in a ligand blot assay (21). This Ab was also able to
immunoprecipitate HARE from isolated LECs. Here we report that mAb-174
also recognizes both HARE species from whole spleen, lymph node, and
liver in Western analysis and performs well in immunolocalization and
confocal microscopy procedures. More notably, mAb-174 also specifically
blocked the uptake and degradation of 125I-HA by isolated
perfused liver. This antibody should, therefore, be a valuable reagent
in future studies of HARE function.
Materials and Media--
125I-HA was
prepared as described (33) using a hexylamine derivative of HA
(oligosaccharides of Mr ~70,000). Male
Sprague-Dawley rats (200 g) were from Charles River Labs. BSA fraction
V was from Intergen. Hanks' balanced salt solution and PBS were
formulated according to the Invitrogen catalog formulations.
Medium 1 was Eagle's basal medium (Invitrogen no. 41500-018)
supplemented with 100 mg/liter succinic acid sodium salt, 75 mg/liter
succinic acid, 2.4 g/liter HEPES, and 0.22g/liter NaHCO3.
Medium 1/BSA was Medium 1 supplemented with 0.1% BSA (w/v).
Collagenase was from Roche Molecular Biochemicals. The preparation of
mouse mAbs against the rat HARE was described (21). Tris, SDS, ammonium
persulfate, N,N'-methylenebisacrylamide, and SDS-PAGE
standards were from Bio-Rad. Rat lymph nodes were a special purchase
from Pel-Freez Biologicals. Unless noted otherwise, other chemicals and
reagents were from Sigma.
Isolation and Culture of LECs--
Rat livers were perfused with
collagenase using a modification of the procedure developed by Seglen
(34) as described previously (35). LECs were isolated using
differential centrifugation and discontinuous Percoll gradients (36).
LECs banding at the 25/50% interface were removed, washed twice with
RPMI 1640 (Invitrogen) containing penicillin/streptomycin (100 units
each) and 2 mM glutamine and suspended at 1.5-2 × 106 cells/ml. The cells were incubated first on a glass
Petri dish for 10 min at room temperature to remove Kupffer cells and
then plated on human fibronectin-coated (50 µg/ml) 24-well tissue
culture plates for endocytosis experiments or on glass coverslips for microscopy. After incubation at 37 °C for 2 h in a 5%
CO2 atmosphere, the cells were washed three times with PBS,
once with RPMI 1640, and then incubated in RPMI 1640 without serum at
37 °C for 1 h if they were to be used immediately or with 2%
heat-inactivated bovine serum if they were to be cultured overnight
before use. The 37 °C pretreatment was performed in all experiments
to allow any endogenous cell surface-bound HA, e.g. from the
serum, to be removed either by internalization or dissociation and washing.
Clearance of 125I-HA by Perfused Liver--
Rat
livers were excised following a standard perfusion protocol (35).
During excision and mounting in a recirculation apparatus, they were
perfused without recirculation with Buffer 1 (142 mM NaCl,
6.7 mM KCl, and 10 mM HEPES, pH 7.4) for 8-10
min at 35 °C. Washed livers were then pre-perfused for 15 min with
recirculation with a recirculation medium (Dulbecco's modified
Eagle's medium, Invitrogen catalog no. 41100, without serum but with
60 mM HEPES, pH 7.4, 0.1% (w/v) BSA, and 50 µg/ml goat
IgG, Sigma catalog no. I-5256). Then the perfusate was switched to 60 ml of fresh recirculation medium containing 125I-HA (0.25 µg/ml in all experiments) and other additions as noted, and the liver
was allowed to take up HA for up to 60 min at 35 °C. Samples (300 µl) of the recirculating perfusate were removed and divided into
50-µl portions for determination of total radioactivity (in
duplicate) or HA degradation products (in triplicate). Competitor HA
(50 µg/ml) or mouse IgG or mAb-174 IgG (5 µg/ml) was added to the
recirculation medium containing the 125I-HA and mixed well
before the recirculating perfusion was begun. The first sample taken
immediately after starting the perfusion was used to determine the
starting values, because there was a substantial (~15%) and
reproducible dilution of the recirculating medium by the residual
buffer in the liver. In experiments with purified IgGs, these were also
included during the 35 °C pre-perfusion treatment at the same
concentration used in the experiment.
Degradation of 125I-HA by Perfused
Liver--
Degradation of 125I-HA was measured by a
cetylpyridinium chloride precipitation assay as described
previously (20). Samples (50 µl in triplicate) of recirculation
medium containing 125I-HA were added to 250 µl of 1 mg/ml
HA in 1.5-ml microfuge tubes. After mixing, 300 µl of 6% (w/v)
cetylpyridinium chloride in distilled water was added, and the tubes
were mixed by vortexing. After 10 min at room temperature, the samples
were centrifuged at 9000 rpm in an Eppendorf model 5417 microcentrifuge
at room temperature for 5 min. A sample (300 µl) of the supernatant
was taken for determination of radioactivity, and the remainder was
removed by aspiration. The tip of the tube containing the precipitate pellet was cut off, put in a Immunocytochemistry--
Lymph node, liver, and spleen tissues
from Sprague-Dawley rats were removed, fixed in 10% neutral buffered
formalin, processed, and paraffin embedded overnight on a Tissue Tek
V.I.P. processor. Tissue sections (5 µm) were collected on charged
slides and dried at 60 °C overnight. The slides were dewaxed three
times for 3 min each with xylene followed by four washes for 3 min each
with alcohol (100%, 95%, 90%, then 70%) and a single 2-min wash in water at room temperature. The endogenous peroxidase activity was
quenched by treatment with 3% hydrogen peroxide for 6 min followed by
two 2-min water washes. The tissue sections on the slides were digested
for 15 min at 37 °C in pre-warmed 0.1 N HCl containing
0.32 mg/ml pepsin followed by a 2-min water wash and a 2-min PBS wash.
The slides were washed with PBS and incubated with the appropriate
primary antibody (~1:500) at room temperature for 60 min. After a 1 min PBS wash, the slides were treated with biotinylated horse
anti-mouse IgG (1:200) for 30 min at room temperature. After another
PBS rinse, the slides were incubated with streptavidin-horseradish peroxidase (1:1000, Jackson Labs) for 30 min, washed once with PBS, and
once with distilled water. Color development was for 5 min with 2.0%
(v/v) aminoethylcarbazine and hydrogen peroxide (ScyTek Laboratories,
Logan Utah) followed by counterstaining with hematoxylin. Slides were
viewed with an Olympus BX-40 light microscope equipped with an Olympus
DP10 digital camera.
General--
Protein content was determined by the method of
Bradford (37) using BSA as standard. Radioiodine was determined using a Packard 5002 To purify and characterize the rat HARE proteins, we developed a
panel of eight mAbs raised against the partially purified 175HARE (21,
30). Surprisingly, all of these mAbs also recognized the 300HARE in
either Western or immuno-purification assays. We now know that this is
because the single 175HARE species and probably the 230-kDa and 250-kDa
subunits of the 300HARE are derived by proteolysis from a larger
precursor (32). Fig. 1 shows the
concentration-dependent blocking of specific uptake of
125I-HA by LECs at 37 °C. Specific internalization was
completely inhibited by 5 µg/ml mAb-174, whereas up to 10 µg/ml of
three other anti-HARE mAbs inhibited only ~15%. Interestingly,
mAb-235 reproducibly inhibited 125I-HA endocytosis by LECs
by about 50%. No further inhibition by the other six mAbs (only
numbers 28, 30, and 467 are shown) or mouse IgG (not shown) occurred
regardless of IgG concentration or duration of treatment.
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
(1,
4)-D-glucuronopyranosyl-
(1, 3). Despite its simple
structure, HA is involved in many cell functions including migration,
differentiation, and phagocytosis (2-6). HA is important in
development (4, 7), wound healing (8, 9), angiogenesis (10, 11), and
tumor growth and metastasis (12, 13). Although previously believed to
be only a structural component in the ECM, HA is now also recognized as
an active cell-signaling molecule. Some cell types show distinct
physiological responses to HA of different sizes. In particular, some
cell types respond physiologically to very small, but not large, HA.
Small HA oligosaccharides containing 14-20 sugars stimulate
angiogenesis by endothelial cells (10, 11, 14), induce gene expression
in activated macrophages (15), and induce NO synthase expression in
sinusoidal LECs and Kupffer cells, but not hepatocytes or stellate
cells (16).
EXPERIMENTAL PROCEDURES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
counter tube, and radioactivity was
determined. Degradation was measured as the time-dependent increase of non-precipitable radioactivity. >80% of the total radioactivity was precipitable at the beginning of the experiment.
counting system. SDS-PAGE was performed according to
the method of Laemmli (38). Western blotting procedures were performed
essentially as described by Burnette (39) with minor modifications
(21).
RESULTS
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
View larger version (26K):
[in a new window]
Fig. 1.
MAb-174 inhibits 125I-HA uptake
by LECs at 37 °C. LECs were plated at 1.9 × 106 cells per well, washed, and pretreated at 37 °C as
described under "Experimental Procedures." The cells were washed
twice with medium, and then medium containing 1 µg/ml
125I-HA and the indicated amount (up to 10 µg/ml) of
mAb-28 ( ), mAb-30 (
), mAb-174 (
), mAb-235 (
), or mAb-467
(
) was added. The dishes were incubated at 37 °C for 60 min, the
medium was then aspirated, and the cells were washed three times with
PBS. The cells were solubilized in 0.3 N NaOH, and
radioactivity and protein content were measured. The values shown are
the average of duplicates (which differed by <5%). The control value,
with no IgG additions, was set to 100%.
Previously, we reported that mAb-174 blocks 125I-HA binding to both HARE species (21) in a ligand blot assay that detects the activity of these proteins after SDS-PAGE, electrotransfer, and renaturation (29). In dose-response experiments, <1 µg/ml, mAb-174 blocked 125I-HA binding to both the affinity-purified 175HARE and 300HARE proteins, but control IgG up to 10 µg/ml had no effect (not shown). Inhibition of binding to the 175HARE was almost complete; ~88% of the specific binding was blocked at 2-10 µg/ml mAb-174. Although the effect of mAb-174 on 125I-HA binding to the 300HARE was identical to that of the 175HARE below 1 µg/ml, inhibition leveled off at ~50% between 1-10 µg/ml.
Despite the ability of mAb-174 to inhibit HA uptake by LECs at 37 °C
or HA binding in the ligand blot assay at room temperature, we
unexpectedly found that mAb-174 did not block 125I-HA
binding to LECs at 4 °C (Fig. 2). To
test the possibility that this inability to inhibit HA binding was due
to an inherent difference in HARE between 37 °C and 4 °C
(e.g. a conformation change), we fixed LECs so that
endocytosis could not occur when the cells were subsequently put at
37 °C. This enabled us to determine whether mAb-174 could block
125I-HA binding per se at 37 °C; this cannot
be assessed if endocytosis occurs simultaneously. The results confirmed
that mAb-174, but none of the other HARE-specific mAbs tested, could
block 125I-HA binding to fixed LECs at 37 °C. We
conclude that HARE likely undergoes a conformation change between
37 °C and 4 °C that does not prevent 125I-HA binding
to LECs but that alters the epitope recognized by mAb-174 so that it
does not block HA binding.
|
We found previously that mAb-174 recognizes the 175HARE and 300HARE
proteins in Western analysis of crude LEC membranes (21). Because we
had not tested any of our mAbs individually using other tissues, we
compared the Western blot reactivity of mAb-174 with the two HARE
species from rat lymph node, spleen, and liver. In all three tissues,
the HARE proteins were visualized by mAb-174 (Fig.
3). Expression of the 175HARE in lymph
node was lower than in the other tissues, although it was detectable
(not shown). In addition to its utility as a ligand-blocking antibody
and for Western analysis, mAb-174 also worked well in a variety of
localization procedures, including indirect fluorescence microscopy
(e.g. the same localization pattern was observed previously
with a mixture of mAbs; Ref. 21) and immunohistochemistry (Fig.
4). Immunohistochemical localization of
HARE using mAb-174 revealed heavy staining of the sinusoidal
endothelial cells in liver, spleen, and lymph node (Fig. 4). The
medullary sinuses of lymph node and the venous sinusoids of spleen
contain large amounts of HARE protein.
|
|
The fact that mAb-174 effectively inhibits HA binding and
uptake by isolated LECs at 37 °C does not necessarily mean that the
antibody would be equally able to inhibit HA clearance by these cells
in intact liver. To answer this question, we tested the ability of
mAb-174 to block 125I-HA uptake and degradation by excised
rat livers that were continuously perfused ex vivo. In these
experiments we used a concentration of mAb-174 (5 µg/ml) that gave
essentially complete inhibition of uptake by LECs. 125I-HA
introduced into the perfusion recirculating medium was readily removed;
~50% was cleared within ~20 min (Fig.
5). In the presence of excess unlabeled
HA, the removal of 125I-HA was completely blocked. Although
control mouse IgG appeared to retard slightly the clearance of
125I-HA, there was no statistically significant difference
between most of the data pairs. In contrast, when mAb-174 was present there were very significant differences in 125I-HA removal
at all times during the perfusion, and the level of residual uptake was
just slightly greater than that in the presence of excess unlabeled HA.
The results confirm that mAb-174 effectively blocks 125I-HA
uptake by LECs in intact perfused liver.
|
This conclusion was further confirmed by demonstrating the effect of
mAb-174 on the appearance of 125I-HA degradation products
in the perfusion medium (Fig. 6).
125I-HA removal and processing by perfused liver was so
efficient that degradation products were detected within minutes. By 30 min, about 25% of the total 125I-HA in the perfusate had
been internalized, degraded, and released back into the medium. Again,
no significant differences were seen in the appearance of degradation
products when mouse IgG was present, although all of the values were
slightly lower. In contrast, mAb-174 substantially reduced the
steady-state rate at which 125I-HA degradation products
were released. The inhibition by unlabeled HA was essentially complete,
indicating that >95% of the observed degradation was mediated by a
specific mechanism.
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
HA is used in many cosmetic and clinical applications, and new HA-containing materials are currently being developed for medical use. For example, HA is used in ophthalmological surgery (40), to treat patients with osteoarthritis in knee and hip joints (41), to prevent adhesions after surgery (42), and as an aerosol to prevent elastase-mediated injury in pulmonary emphysema (43). Because of the wide range of medical uses for HA, it is very important to understand how HA turnover and clearance are regulated. The present study was undertaken as an initial effort to define the role of HARE in this process.
After LECs were found to contain an endocytic HA receptor (23, 44), others demonstrated that isolated perfused rat liver can remove circulating HA (24, 25, 45), presumably mediated by this HA receptor. The present results verify that this assumption was correct. The quantitative inhibition of 125I-HA uptake and degradation by mAb-174 demonstrates that HARE mediates at least 90% of the liver's clearance ability. In this study, we did not perform an extensive dose response analysis because of the larger number of animals and amount of purified IgG required. Accordingly, we did not necessarily use an optimum dose of mAb-174 for blocking HARE in perfused liver, although the mAb-174 concentration used, 5 µg/ml, gave maximum inhibition of 125I-HA internalization by LECs (Fig. 1).
Using a mixture of anti-HARE mAbs, we found that the small and large rat HARE proteins are highly expressed in the sinusoids of liver, the venous sinuses of the red pulp in spleen, and the medullary sinuses in lymph nodes (21). This distribution was also seen using only mAb-174 (Fig. 4). HARE was not detectable by Western or immuno-cytochemical analysis in brain, lung, heart, muscle, kidney, or intestine. Abundant expression of HARE in the sinusoids of liver and lymphatic tissues is ideal for keeping the systemic HA levels low. Banerji et al. (46) discovered a lymph-specific homologue of CD44, designated LYVE-1, which binds HA and is localized to the luminal face of vessel walls in the lymphatic system. It is not present on blood vessels. Preliminary studies indicate that LYVE-1 and HARE have different and non-overlapping distributions in lymph node and spleen and are not co-localized even within isolated rat LECs.
The rat 175HARE is a 1431-amino acid (156,393 Da) type I membrane protein with a large NH2-terminal extracellular domain (~1323 amino acids), one transmembrane domain (~20 amino acids), and a small COOH-terminal cytoplasmic domain (~88 amino acids). The native protein contains ~25 kDa of N-linked oligosaccharides (30). The recombinant rat 175HARE protein, expressed in SK-Hep-1 cells, is a functional HA receptor able to mediate the specific and continuous endocytosis of 125I-HA through the clathrin coated pit pathway (32). Because the 175HARE is functional in the absence of the 300HARE complex, it is not necessary for these two HARE species to be present in the same cell in order to create a specific, endocytically competent HA receptor. Therefore, the 175HARE and 300HARE are independent isoreceptors for HA. Our present results show that mAb-174 blocks HA binding and endocytosis by both HARE species in intact liver.
A 70-kg person contains about 15 g of HA and turns over about 5 g, or one-third of their total HA, per day (47). About 50% of the total HA is in skin and has a metabolic half-life of <1.5 days (48). Lymph nodes and liver are the major clearance sites for the systemic removal and degradation of HA (2). HA injected intravenously in mammals is rapidly removed from the blood by liver (2, 45, 47), whereas HA introduced into lymph is degraded first by lymph nodes, and the remainder is then removed by the liver. The present model of mammalian HA turnover is that large native HA (~107 Da) in ECMs throughout the body is partially degraded to fragments of ~106 Da that are released into lymphatic vessels and then flow to lymph nodes where ~85% of the total HA is removed and degraded. The remaining HA (~15%) that exits the lymph nodes is ~105 Da and, after entering the blood, is removed by the LECs of liver.
Despite the generation of up to 5 g of HA degradation products per day, the HA clearance systems utilizing HARE in lymph nodes and liver keep the normal steady-state HA concentration in blood very low (i.e. 10-100 ng/ml). Clearance of circulating HA from blood is important for normal health, because blood viscosity could increase to levels that might impair the microcirculation if the concentration of large HA (i.e. >106 Da) increased. Blood HA levels could rise due either to greater HA production or turnover in the body, exceeding the capacity of LECs to remove it, or to a compromised function of HARE in lymph nodes or liver. Elevated blood HA levels are found in several diseases, although in most cases the cause of this elevation is not known. Elevated serum HA is now used as a diagnostic indicator of liver failure. For example, elevated serum HA is a marker for liver fibrosis in hepatitis C virus-associated chronic liver disease (49). Increased ascitic levels of HA occur in liver cirrhosis due to increased HA synthesis by peritoneal cells and decreased uptake by LECs (50). Elevated blood HA levels have also been reported in a variety of other diseases such as rheumatoid arthritis (51), psoriasis (52), and in some cancers (53). Blood HA levels can also predict the functionality of LECs and the likely success of liver transplants (26, 27).
In summary, mAb-174 specifically inhibits the HA-binding and
HA-clearance activities of both the small and large HARE proteins in
isolated rat LECs and in whole perfused liver. This effective activity-blocking antibody is particularly versatile, because it is
also functional in Western analysis and a variety of other immuno-procedures and should be useful in future studies to understand the physiological functions of HARE.
![]() |
FOOTNOTES |
---|
* This research was supported by NIGMS, National Institutes of Health Grant GM35978.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
¶ Present address: Dept. of Pathology, St. Joseph's Hospital, 69 Exchange St., St. Paul, MN 55102.
To whom correspondence should be addressed. Tel.:
405-271-1288; Fax: 405-271-3092; E-mail: paul-weigel@ouhsc.edu.
Published, JBC Papers in Press, January 6, 2003, DOI 10.1074/jbc.M211462200
![]() |
ABBREVIATIONS |
---|
The abbreviations used are: HA, hyaluronic acid, hyaluronate, or hyaluronan; BSA, bovine serum albumin; ECM, extracellular matrix; HARE, HA receptor for endocytosis; 175HARE, 175-kDa HARE; 300HARE, 300-kDa HARE; ICAM-1, intercellular adhesion molecule-1 (also designated CD54); LEC, sinusoidal liver endothelial cell; LYVE, lymphatic vessel endothelial HA receptor; mAb, monoclonal antibody; PBS, phosphate buffered saline; RHAMM, receptor for HA-mediated motility.
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1. |
Meyer, K.,
and Palmer, J. W.
(1934)
J. Biol. Chem.
107,
629-634 |
2. |
Laurent, T. C.,
and Fraser, J. R. E.
(1992)
FASEB J.
6,
2397-2404 |
3. |
Knudson, C. B.,
and Knudson, W.
(1993)
FASEB J.
7,
1233-1241 |
4. | Toole, B. P. (1997) J. Intern. Med. 242, 35-40[Medline] [Order article via Infotrieve] |
5. | Abatangelo, G., and Weigel, P. H. (eds) (2000) New Frontiers in Medical Sciences: Redefining Hyaluronan , Elsevier Science Publishers B. V., Amsterdam |
6. | Turley, E. A. (1992) Cancer Metastasis Rev. 11, 1-3[Medline] [Order article via Infotrieve] |
7. |
Gakunga, P.,
Frost, G.,
Shuster, S.,
Cunha, G.,
Formby, B.,
and Stern, R.
(1997)
Development
124,
3987-3997 |
8. | Burd, D. A. R., Greco, R. M., Regauer, S., Longaker, M. T., Siebert, J. W., and Garg, H. G. (1991) Br. J. Plast. Surg. 44, 579-584[Medline] [Order article via Infotrieve] |
9. | Chen, W. Y., and Abatangelo, G. (1999) Wound Repair Regen. 7, 79-89[CrossRef][Medline] [Order article via Infotrieve] |
10. | West, D. C., Hampson, I. N., Arnold, F., and Kumar, S. (1985) Science 14, 1324-1326 |
11. | Deed, R., Rooney, P., Kumar, P., Norton, J. D., Smith, J., Freemont, A. J., and Kumar, S. (1997) Int. J. Cancer 71, 251-256[CrossRef][Medline] [Order article via Infotrieve] |
12. | Csoka, T. B., Frost, G. I., and Stern, R. (1997) Invasion Metastasis 17, 297-311[Medline] [Order article via Infotrieve] |
13. | Delpech, B., Girard, N., Bertrand, P., Courel, M. N., Chauzy, C., and Delpech, A. (1997) J. Intern. Med. 242, 41-48[Medline] [Order article via Infotrieve] |
14. | Rahmanian, M., Pertoft, H., Kanda, S., Christofferson, R., Claesson-Welsh, L., and Heldin, P. (1997) Exp. Cell Res. 237, 223-230[CrossRef][Medline] [Order article via Infotrieve] |
15. | Horton, M. R., Olman, M. A., Bao, C., White, K. E., Choi, A. M., Chin, B. Y., Noble, P. W., and Lowenstein, C. (2002) Am. J. Physiol. Lung Cell. Mol. Physiol. 279, L707-L715 |
16. | Rockey, D. C., Chung, J. J., Mckee, C. M., and Noble, P. W. (1998) Hepatology 27, 86-92[Medline] [Order article via Infotrieve] |
17. | Weigel, P. H. (1992) Mechanisms and Control of Glycoconjugate Turnover. Glycoconjugates: Composition, Structure, and Function , pp. 421-497, Marcel Dekker Inc., New York |
18. |
Raja, R. H.,
McGary, C. T.,
and Weigel, P. H.
(1988)
J. Biol. Chem.
263,
16661-16668 |
19. | McGary, C. T., Raja, R. H., and Weigel, PH. (1989) Biochem. J. 257, 875-884[Medline] [Order article via Infotrieve] |
20. | McGary, C. T., Yannariello-Brown, J., Kim, D. W., Stinson, T. C., and Weigel, P. H. (1993) Hepatology 18, 1465-1476[Medline] [Order article via Infotrieve] |
21. |
Zhou, B.,
Weigel, J. A.,
Fauss, L.,
and Weigel, P. H.
(2000)
J. Biol. Chem.
275,
37733-37741 |
22. | Laurent, T. C., Fraser, J. R., Pertoft, H., and Smedsrod, B. (1986) Biochem. J. 234, 653-658[Medline] [Order article via Infotrieve] |
23. | Weigel, P. H., and Yik, J. H. N. (2002) Biochim. Biophys. Acta 1572, 341-363[Medline] [Order article via Infotrieve] |
24. | Deaciuc, I. V., Bagby, G. J., and Spitzer, J. J. (1993) Biochem. Pharmacol. 46, 671-675[CrossRef][Medline] [Order article via Infotrieve] |
25. | Deaciuc, I. V., Bagby, G. J., Lang, C. H., and Spitzer, J. J. (1993) Hepatology 17, 266-272[Medline] [Order article via Infotrieve] |
26. | Itasaka, H., Suehiro, T., Wakiyama, S., Yanaga, K., Shimada, M., and Sugimachi, K. (1995) J. Surg. Res. 59, 589-595[CrossRef][Medline] [Order article via Infotrieve] |
27. | Rao, P. N., Bronsther, O. L., Pinna, A. D., Snyder, J. T., Cowan, S., Sankey, S., Kramer, D., Takaya, S., and Starzl, T. (1996) Liver 16, 48-54[Medline] [Order article via Infotrieve] |
28. | Yannariello-Brown, J., and Weigel, P. H. (1992) Biochemistry 31, 576-584[Medline] [Order article via Infotrieve] |
29. | Yannariello-Brown, J., Zhou, B., Ritchie, D., Oka, J. A., and Weigel, P. H. (1996) Biochem. Biophys. Res. Commun 218, 314-319[CrossRef][Medline] [Order article via Infotrieve] |
30. |
Zhou, B.,
Oka, J. A.,
and Weigel, P. H.
(1999)
J. Biol. Chem.
274,
33831-33834 |
31. | Zhou, B., McGary, C. T., Weigel, J. A., Saxena, A., and Weigel, P. H. (2003) Glycobiology, in press |
32. |
Zhou, B.,
Weigel, J. A.,
Saxena, A.,
and Weigel, P. H.
(2002)
Mol. Biol. Cell
13,
2853-2868 |
33. | Raja, R. H., LeBoeuf, R. D., Stone, G. W., and Weigel, P. H. (1984) Anal. Biochem. 139, 168-177[Medline] [Order article via Infotrieve] |
34. | Seglen, P. O. (1976) Methods Cell Biol. 13, 29-83[Medline] [Order article via Infotrieve] |
35. |
Clarke, B. L.,
Oka, J. A.,
and Weigel, P. H.
(1987)
J. Biol. Chem.
262,
17384-17392 |
36. | Smedsrod, B., Pertoft, H., Eggersten, G., and Sundstrom, C. (1985) Cell Tissue Res. 241, 639-649[Medline] [Order article via Infotrieve] |
37. | Bradford, M. M. (1976) Anal. Biochem. 72, 248-254[CrossRef][Medline] [Order article via Infotrieve] |
38. | Laemmli, U. K. (1970) Nature 227, 680-685[Medline] [Order article via Infotrieve] |
39. | Burnette, W. N. (1981) Anal. Biochem. 112, 195-203[Medline] [Order article via Infotrieve] |
40. | Goa, K. L., and Benfield, P. (1994) Drugs 47, 536-566[Medline] [Order article via Infotrieve] |
41. | Manek, N. J., and Lane, N. E. (2000) Am. Fam. Physician 61, 1795-1804[Medline] [Order article via Infotrieve] |
42. | Panay, N., and Lower, A. M. (1999) Curr. Opin. Obstet. Gynecol. 11, 379-385[CrossRef][Medline] [Order article via Infotrieve] |
43. | Cantor, J. O., Cerreta, J. M., Armand, G., and Turino, G. M (1998) Proc. Soc. Exp. Biol. Med. 217, 471-475[Abstract] |
44. | Mellman, I. (1996) Annu. Rev. Cell Dev. Biol. 12, 575-625[CrossRef][Medline] [Order article via Infotrieve] |
45. | Fraser, J. R., Appelgren, L. E., and Laurent, T. C. (1983) Cell Tissue Res. 233, 285-293[Medline] [Order article via Infotrieve] |
46. | Banerji, S., Day, A. J., Kahmann, J. D., and Jackson, D. G. (1998) Protein Expression Purif. 14, 371-381[CrossRef][Medline] [Order article via Infotrieve] |
47. | Laurent, T. C., and Fraser, J. R. E. (1991) in Degradation of Bioactive Substances: Physiology and Pathophysiology (Henriksen, J. H., ed) , pp. 249-265, CRC Press, Boca Raton, FL |
48. | Tammi, R., Saamanen, A. M., Maibach, H. I., and Tammi, M. (1991) J. Invest. Dermatol. 97, 126-130[Abstract] |
49. | Fraser, J. R., Laurent, T. C., Pertoft, H., and Baxter, E. (1981) Biochem. J. 200, 415-424[Medline] [Order article via Infotrieve] |
50. | Yamada, M., Fukuda, Y., Nakano, I., Katano, Y., Takamatsu, J., and Hayakawa, T. (1998) Acta Haematol. 99, 212-216[CrossRef][Medline] [Order article via Infotrieve] |
51. | Manicourt, D. H., Poilvache, P., Nzeusseu, A., van Egeren, A., Devogelaer, J. P., Lenz, M. E., and Thonar, E, J. (1999) Arthritis Rheum. 42, 1861-1869[CrossRef][Medline] [Order article via Infotrieve] |
52. | Lundin, A., Engstrom-Laurent, A., Hallgren, R., and Michaelsson, G. (1985) Br. J. Dermatol. 112, 663-671[Medline] [Order article via Infotrieve] |
53. | Thylen, A., Wallin, J., and Martensson, G. (1999) Cancer 86, 2000-2005[CrossRef][Medline] [Order article via Infotrieve] |