On the adsorption of hyaluronan and ICAM-1 to modified hydrophobic resins

McCourt, Peter; Gustafson, Stefan

doi:10.1016/s1357-2725(97)00058-7

Cited by 26 publications

(19 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, not all HA-binding proteins contain a link domain, namely, RHAMM, Cdc37, inter-␣-trypsin inhibitor, plasma HA-binding protein, fibroblast HA-binding protein (reviewed by Day, 1999), and intracellular hyaluronate-binding protein (Hofmann et al, 1998). The best characterized HA cell surface receptors are CD44, RHAMM, ICAM-1, and LYVE-1 (Entwistle et al, 1996;Banerji et al, 1999), although there is some controversy as to whether ICAM-1 is a genuine HA receptor (McCourt and Gustafson, 1997).…”

Section: Discussionmentioning

confidence: 99%

“…HA-EDA-Sepharose 4B (McCourt and Gustafson, 1997) was kindly provided by Dr. Paraskevi Heldin (Uppsala University, Sweden). Unsubstituted Sepharose 4B was used as a control in the binding reactions.…”

Section: Fc Fusion Protein Binding To Ha Immobilized To Sepharosementioning

confidence: 99%

“…HA is a linear polysaccharide, which can be immobilized, for example, to EDA-Sepharose (McCourt and Gustafson, 1997). Because HA-binding proteins can be purified by affinity chromatography on immobilized HA (Tengblad, 1979), we incubated layilin-IgG as well as control IgG with HA-Sepharose beads.…”

Section: Ha Is a Ligand For Layilinmentioning

confidence: 99%

See 2 more Smart Citations

Layilin, a Novel Integral Membrane Protein, Is a Hyaluronan Receptor

Bono

Rubin

Higgins

et al. 2001

MBoC

131

View full text Add to dashboard Cite

The actin cytoskeleton plays a significant role in changes of cell shape and motility, and interactions between the actin filaments and the cell membrane are crucial for a variety of cellular processes. Several adaptor proteins, including talin, maintain the cytoskeleton-membrane linkage by binding to integral membrane proteins and to the cytoskeleton. Layilin, a recently characterized transmembrane protein with homology to C-type lectins, is a membrane-binding site for talin in peripheral ruffles of spreading cells. To facilitate studies of layilin's function, we have generated a layilin-Fc fusion protein comprising the extracellular part of layilin joined to human immunoglobulin G heavy chain and used this chimera to identify layilin ligands. Here, we demonstrate that layilin-Fc fusion protein binds to hyaluronan immobilized to Sepharose. Microtiter platebinding assays, coprecipitation experiments, and staining of sections predigested with different glycosaminoglycan-degrading enzymes and cell adhesion assays all revealed that layilin binds specifically to hyaluronan but not to other tested glycosaminoglycans. Layilin's ability to bind hyaluronan, a ubiquitous extracellular matrix component, reveals an interesting parallel between layilin and CD44, because both can bind to cytoskeleton-membrane linker proteins through their cytoplasmic domains and to hyaluronan through their extracellular domains. This parallelism suggests a role for layilin in cell adhesion and motility. INTRODUCTIONCell shape and movement form the basis for several biological phenomena such as morphogenesis, invasion, and metastasis. The actin cytoskeleton is a major determinant of changes in cell shape, and interactions between actin filaments and the cell membrane are essential for cell adhesion, spreading, and migration, as well as for signal transduction (Hall, 1998;Schoenwaelder and Burridge, 1999). Several molecules have been recognized as linkers between the actin cytoskeleton and the cell membrane. For example, members of the band 4.1/ERM superfamily (band 4.1, talin, ezrin, radixin, moesin, and merlin, the product of the neurofibromatosis type 2 tumor suppressor gene) are characterized by the presence of a conserved N-terminal membrane-binding domain, which is able to bind to the cytoplasmic region of several transmembrane receptors, including CD44, CD43, ICAM-2, and ICAM-3 (Sainio et al., 1997;Heiska et al., 1998;Yonemura et al., 1998). The C-terminal regions of ERM proteins are thought to bind to actin filaments, thereby linking these transmembrane receptors to the cytoskeleton (reviewed by Mangeat et al., 1999).Layilin is a recently cloned ϳ55-kDa membrane-binding partner for talin (Borowsky and Hynes, 1998) that is widely expressed in different cell types and tissue extracts. It is found in peripheral ruffles of spreading cells and is recruited to membrane ruffles in cells induced to migrate in in vitro wounding experiments. Layilin colocalizes with talin in ruffles and binds to talin's ϳ50-kDa head domain (amino acids 280 -435)...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Fc Fusion Protein Binding To Ha Immobilized To Sepharosementioning

confidence: 99%

See 1 more Smart Citation

Layilin, a Novel Integral Membrane Protein, Is a Hyaluronan Receptor

Bono

Rubin

Higgins

et al. 2001

MBoC

131

View full text Add to dashboard Cite

show abstract

“…Earlier reports misidentified this LEC HAR as ICAM-1 (11,12), also known as CD54, which is a 90-kDa protein. This finding was later recognized as an artifact in that ICAM-1 bound nonspecifically to the HA affinity resin employed (13). In two previous studies, one using a photoaffinity derivative of HA (14) and the other using a novel ligand blot assay with 125 I-HA (15), we identified two specific HA-binding proteins in isolated rat LECs at 175 and ϳ300 kDa.…”

Section: Hamentioning

confidence: 93%

Purification and Subunit Characterization of the Rat Liver Endocytic Hyaluronan Receptor

Zhou

Oka

Singh

et al. 1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

The endocytic hyaluronan (HA) receptor of liver sinusoidal endothelial cells (LECs) is responsible for the clearance of HA and other glycosaminoglycans from the circulation in mammals. We report here for the first time the purification of this liver HA receptor. Using lectin and immuno-affinity chromatography, two HA receptor species were purified from detergent-solubilized membranes prepared from purified rat LECs. In nonreducing SDS-polyacrylamide gel electrophoresis (PAGE), these two proteins migrated at 175-and ϳ300 kDa corresponding to the two species previously identified by photoaffinity labeling of live cells as the HA receptor (Yannariello-Brown, J., Frost, S. J., and Weigel, P. H. (1992) J. Biol. Chem. 267, 20451-20456). These two proteins co-purify in a molar ratio of 2:1 (175:300), and both proteins are active, able to bind HA after SDS-PAGE, electrotransfer, and renaturation. After reduction, the 175-kDa protein migrates as a ϳ185-kDa protein and is not able to bind HA. The 300-kDa HA receptor is a complex of three disulfide-bonded subunits that migrate in reducing SDS-PAGE at ϳ260, 230, and 97 kDa. These proteins designated, respectively, the ␣, ␤, and ␥ subunits are present in a molar ratio of 1:1:1 and are also unable to bind HA when reduced. The 175-kDa protein and all three subunits of the 300-kDa species contain N-linked oligosaccharides, as indicated by increased migration in SDS-PAGE after treatment with N-glycosidase F. Both of the deglycosylated, nonreduced HA receptor proteins still bind HA.HA 1 is an important extracellular matrix component of all tissues and plays a key role in development, cell proliferation, cell adhesion, recognition, morphogenesis, differentiation, and inflammation (1-4). The daily total body turnover of HA in humans is at least 1 g/day (4). HA degradation and removal in the body occurs via two clearance systems (3): one is in the lymphatic system, which accounts for ϳ85% of the HA turnover, and another system is hepatic, accounting for ϳ15% of the total body HA turnover. HA in tissues throughout the body is continuously synthesized and degraded. Very large HA molecules (ϳ10 7 Da) are partially degraded to large fragments (ϳ10 6 Da) that are then released from the matrix and flow with the lymph to lymph nodes. The majority of HA (ϳ85%) is completely degraded in the lymph nodes by unknown mechanisms and the remaining HA (ϳ15%) that passes through the nodes finally enters the blood. Clearance of this circulating HA is presumably important for normal health (3, 4). Elevated serum HA levels are found in several disease conditions such as liver cirrhosis, rheumatoid arthritis, psoriasis, scleroderma, and some cancers (5-7).LECs have a very active recycling, endocytic receptor that removes HA and other glycosaminoglycans, such as chondroitin sulfate, from the circulation (3, 8 -10). Earlier reports misidentified this LEC HAR as ICAM-1 (11, 12), also known as CD54, which is a 90-kDa protein. This finding was later recognized as an artifact in that ICAM-1 bound nonspecifica...

show abstract

“…These workers attempted to purify the HA receptor without the use of an assay to measure HA binding activity. The claim that the HA receptor had been purified was subsequently acknowledged to be an artifact due to the nonspecific binding of ICAM-1 to the HA affinity resin (17,18). In any case, since ICAM-1 is not a coated pit targeted endocytic receptor, it could not be the true HA receptor in LECs.…”

Section: Hamentioning

confidence: 99%

Identification of the Hyaluronan Receptor for Endocytosis (HARE)

Zhou

Weigel

Fauss

et al. 2000

Journal of Biological Chemistry

239

222

View full text Add to dashboard Cite

Rat liver sinusoidal endothelial cells (LECs) express two hyaluronan (HA) receptors, of 175 and 300 kDa, responsible for the endocytic clearance of HA. We have characterized eight monoclonal antibodies (mAbs) raised against the 175-kDa HA receptor partially purified from rat LECs. These mAbs also cross-react with the 300-kDa HA receptor. The 175-kDa HA receptor is a single protein, whereas the 300-kDa species contains three subunits, ␣, ␤, and ␥ at 260, 230, and 97 kDa, respectively (Zhou, B., Oka, J. A., and Weigel, P. H. (1999) J. Biol. Chem. 274, 33831-33834). The 97-kDa subunit was not recognized by any of the mAbs in Western blots. Based on their cross-reactivity with these mAbs, the 175-, 230-, and 260-kDa proteins appear to be related. Two of the mAbs inhibit 125 I-HA binding and endocytosis by LECs at 37°C. All of these results confirm that the mAbs recognize the bone fide LEC HA receptor. Indirect immunofluoresence shows high protein expression in liver sinusoids, the venous sinuses of the red pulp in spleen, and the medullary sinuses of lymph nodes. Because the tissue distribution for this endocytic HA receptor is not unique to liver, we propose the name HARE (HA receptor for endocytosis). HA1 is an important and often abundant extracellular matrix component of all tissues, in particular cartilage, skin, and vitreous humor (1). HA plays a key role in development, morphogenesis, and differentiation, in cell adhesion and proliferation, and in inflammation and wound healing (1-4). In humans the total body turnover of HA is several grams per day (1). Although local turnover of HA occurs in avascular tissues, particularly cartilage (5, 6), two major clearance systems are responsible for HA degradation and removal in the body (4). The first is the lymphatic system, which accounts for ϳ85% of the HA turnover, and the second is in the liver, which accounts for the other ϳ15% of the total body HA turnover. Throughout the body, HA is continuously synthesized and degraded in almost all tissues. At the same time, chondroitin sulfate and other glycosaminoglycans are also released from the cleavage of proteoglycans, especially aggregating proteoglycans associated with HA. Large native HA molecules (ϳ10 7 Da) are partially degraded to large fragments (ϳ10 6 Da) that are released from the matrix and enter the lymphatic system, flowing to lymph nodes.The lymph nodes completely degrade the majority of HA (ϳ85%) by unknown mechanisms. Neither the responsible cell type, the receptor involved, nor the location in lymph nodes at which HA uptake and degradation occurs has been determined. The remaining HA (ϳ15%) that escapes degradation in the lymph nodes ultimately flows into the blood at the thoracic duct. Since HA is an exceptionally viscous polysaccharide in solution, it would be deleterious for the blood concentration of HA, even at relatively low molecular weight, to increase. Clearance of this circulating HA and the other glycosaminoglycan degradation fragments is presumably important for normal health (1, 4). Elevate...

show abstract

On the adsorption of hyaluronan and ICAM-1 to modified hydrophobic resins

Cited by 26 publications

References 22 publications

Layilin, a Novel Integral Membrane Protein, Is a Hyaluronan Receptor

Layilin, a Novel Integral Membrane Protein, Is a Hyaluronan Receptor

Purification and Subunit Characterization of the Rat Liver Endocytic Hyaluronan Receptor

Identification of the Hyaluronan Receptor for Endocytosis (HARE)

Contact Info

Product

Resources

About