Identification of the zinc-dependent endothelial cell binding protein for high molecular weight kininogen and factor XII: Identity with the receptor that binds to the globular ''heads'' of C1q (gC1q-R)

被引:209
作者
Joseph, K
Ghebrehiwet, B
Peerschke, EIB
Reid, KBM
Kaplan, AP
机构
[1] SUNY STONY BROOK,HLTH SCI CTR,DEPT MED,STONY BROOK,NY 11794
[2] SUNY STONY BROOK,DEPT PATHOL,STONY BROOK,NY 11794
[3] UNIV OXFORD,DEPT BIOCHEM,MRC,IMMUNOCHEM UNIT,OXFORD OX1 3QU,ENGLAND
关键词
D O I
10.1073/pnas.93.16.8552
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
High molecular weight kininogen (HK) and factor XII are known to bind to human umbilical vein endothelial cells (HUVEC) in a zinc-dependent and saturable manner indicating that HUVEC express specific binding site(s) for those proteins. However, identification and immunochemical characterization of the putative receptor site(s) has not been previously accomplished. In this report, we have identified a cell surface glycoprotein that is a likely candidate for the HK binding site on HUVECs. When solubilized HUVEC membranes were subjected to an HK-affinity column in the presence or absence of 50 mu M ZnCl2 and the bound membrane proteins eluted, a single major protein peak was obtained only in the presence of zinc. SDS/PAGE analysis and silver staining of the protein peak revealed this protein to be 33 kDa and partial sequence analysis matched the NH2 terminus of gC1q-R, a membrane glycoprotein that binds to the globular ''heads'' of C1q. Two other minor proteins of approximate to 70 kDa and 45 kDa were also obtained. Upon analysis by Western blotting, the 33-kDa band was found to react with several monoclonal antibodies (mAbs) recognizing different epitopes on gC1q-R. Ligand and dot blot analyses revealed zinc-dependent binding of biotinylated HK as well as biotinylated factor XII to the isolated 33-kDa HUVEC molecule as well as recombinant gC1q-R. In addition, binding of I-125-HK to HUVEC cells was inhibited by selected monoclonal anti-gC1q-R antibodies. C1q, however, did not inhibit I-125-HK binding to HUVEC nor did those monoclonals known to inhibit C1q binding to gC1q-R. Taken together, the data suggest that HK (and factor XII) bind to HUVECs via a 33-kDa cell surface glycoprotein that appears to be identical to gC1q-R but interact with a site on gC1q-R distinct from that which binds C1q.
引用
收藏
页码:8552 / 8557
页数:6
相关论文
共 37 条
[1]  
BERRETTINI M, 1992, J BIOL CHEM, V267, P19833
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]  
DELACADENA RA, 1994, THROMB HAEMOSTASIS, V72, P125
[4]   IDENTIFICATION OF A GC1Q-BINDING PROTEIN (GC1Q-R) ON THE SURFACE OF HUMAN NEUTROPHILS - SUBCELLULAR-LOCALIZATION AND BINDING-PROPERTIES IN COMPARISON WITH THE CC1Q-R [J].
EGGLETON, P ;
GHEBREHIWET, B ;
SASTRY, KN ;
COBURN, JP ;
ZANER, KS ;
REID, KBM ;
TAUBER, AI .
JOURNAL OF CLINICAL INVESTIGATION, 1995, 95 (04) :1569-1578
[5]  
GHEBREHIWET B, 1995, J IMMUNOL, V155, P2614
[6]   ISOLATION, CDNA CLONING, AND OVEREXPRESSION OF A 33-KD CELL-SURFACE GLYCOPROTEIN THAT BINDS TO THE GLOBULAR HEADS OF C1Q [J].
GHEBREHIWET, B ;
LIM, BL ;
PEERSCHKE, EIB ;
WILLIS, AC ;
REID, KBM .
JOURNAL OF EXPERIMENTAL MEDICINE, 1994, 179 (06) :1809-1821
[7]  
Ghebrehiwet B, 1989, Behring Inst Mitt, P204
[8]   RECEPTORS FOR HIGH MOLECULAR-WEIGHT KININOGEN ON STIMULATED WASHED HUMAN-PLATELETS [J].
GREENGARD, JS ;
GRIFFIN, JH .
BIOCHEMISTRY, 1984, 23 (26) :6863-6869
[9]   HIGH-MOLECULAR-WEIGHT KININOGEN BINDS TO UNSTIMULATED PLATELETS [J].
GUSTAFSON, EJ ;
SCHUTSKY, D ;
KNIGHT, LC ;
SCHMAIER, AH .
JOURNAL OF CLINICAL INVESTIGATION, 1986, 78 (01) :310-318
[10]   HUMAN-NEUTROPHILS CONTAIN AND BIND HIGH MOLECULAR-WEIGHT KININOGEN [J].
GUSTAFSON, EJ ;
SCHMAIER, AH ;
WACHTFOGEL, YT ;
KAUFMAN, N ;
KUCICH, U ;
COLMAN, RW .
JOURNAL OF CLINICAL INVESTIGATION, 1989, 84 (01) :28-35