Epitope mapping of inhibitory antibodies against platelet glycoprotein Ibα reveals interaction between the leucine-rich repeat N-terminal and C-terminal flanking domains of glycoprotein Ibα

被引:67
作者
Cauwenberghs, N
Vanhoorelbeke, K
Vauterin, S
Westra, DF
Rome, G
Huizinga, EG
Lopez, JA
Berndt, MC
Harsfalvi, J
Deckmyn, H
机构
[1] Katholieke Univ Leuven, Interdisciplinary Res Ctr, Lab Thrombosis Res, B-8500 Kortrijk, Belgium
[2] Univ Utrecht Hosp, Dept Haematol, Utrecht, Netherlands
[3] Baylor Coll Med, Thrombosis Res Sect, Houston, TX 77030 USA
[4] Baker Med Res Inst, Melbourne, Vic, Australia
[5] Univ Debrecen, Sch Med, Dept Clin Biochem & Mol Pathol, Debrecen, Hungary
关键词
D O I
10.1182/blood.V98.3.652
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The interaction of von Willebrand factor (vWF) with the platelet receptor glycoprotein Ib alpha (GPIb alpha) is important for platelet adhesion at high shear stress. Two functionally important antigenic areas within GPIb alpha were identified through the characterization of 5 new inhibitory anti-GPIb monoclonal antibodies (mAbs). The binding sites of 3 of these anti-GPIb mAbs, which were intercompeting and potently inhibiting shear stress-induced binding of vWF, were mapped within the N-terminal amino acid (aa) 1-59 area by the use of canine-human chimeras. These antibodies, however, had little or no effect (approximately 40% inhibition) on the binding of vWF induced by either botrocetin or ristocetin. On the other hand, the anti-GPIb mAbs 24G10 and 6B4, which blocked GPIb-vWF binding under all conditions examined, bound to 2 different regions of GPIb alpha, aa 1-81 and aa 201-268, respectively. The epitope for 6B4 was further narrowed by phage display revealing 2 sets of peptide sequences aligning within aa 259-262 and aa 230-242. In the latter region of GPIb alpha, the gain-of-function platelet-type von Willebrand disease (PT-vWD) mutations have been identified. Alignment was partially confirmed because the binding of 6B4 to recombinant GPIb alpha fragments carrying either one of the PT-vWD mutations was considerably impaired but not completely abolished. In contrast, mAb 24G10 bound more strongly to mutant PT-vWD GPIb alpha. However, although 24G10 competed with 6B4 for binding to platelets, it bound to an epitope within aa 1-81 of GPIb alpha. In conclusion, 2 functionally important areas within GPlb alpha were identified: one localized within the leucine-rich repeat N-terminal aa 1-59 area and one composed of residues aa 1-81 In close contact with aa 201-268. Moreover, further support is provided for the existence of an intramolecular interaction between the N-terminal flanking (aa 1-81) and C-terminal flanking (aa 201-268) regions.
引用
收藏
页码:652 / 660
页数:9
相关论文
共 38 条
[1]   Epitope mapping by screening of phage display libraries of a monoclonal antibody directed against the receptor binding domain of human alpha 2-macroglobulin [J].
Birkenmeier, G ;
Osman, AA ;
Kopperschlager, G ;
Mothes, T .
FEBS LETTERS, 1997, 416 (02) :193-196
[2]   Probing the basis of antibody reactivity with a panel of constrained peptide libraries displayed by filamentous phage [J].
Bonnycastle, LLC ;
Mehroke, JS ;
Rashed, M ;
Gong, X ;
Scott, JK .
JOURNAL OF MOLECULAR BIOLOGY, 1996, 258 (05) :747-762
[3]  
Cauwenberghs N, 2000, HAEMOSTASIS, V30, P139
[4]  
Clemetson KJ, 1997, THROMB HAEMOSTASIS, V78, P266
[5]  
Depraetere H, 1998, BLOOD, V91, P3792
[6]   Ristocetin-dependent, but not botrocetin-dependent, binding of von Willebrand factor to the platelet glycoprotein Ib-IX-V complex correlates with shear-dependent interactions [J].
Dong, JF ;
Berndt, MC ;
Schade, A ;
McIntire, LV ;
Andrews, RK ;
López, JA .
BLOOD, 2001, 97 (01) :162-168
[7]   Crossreactivity of mimotopes and peptide homologues of a sequential epitope with a monoclonal antibody does not predict crossreactive immunogenicity [J].
El Kasmi, KC ;
Deroo, S ;
Theisen, DM ;
Brons, NHC ;
Muller, CP .
VACCINE, 1999, 18 (3-4) :284-290
[8]   BIOSPECIFIC INTERACTION ANALYSIS USING SURFACE-PLASMON RESONANCE DETECTION APPLIED TO KINETIC, BINDING-SITE AND CONCENTRATION ANALYSIS [J].
FAGERSTAM, LG ;
FROSTELLKARLSSON, A ;
KARLSSON, R ;
PERSSON, B ;
RONNBERG, I .
JOURNAL OF CHROMATOGRAPHY, 1992, 597 (1-2) :397-410
[9]   MIMICKING OF DISCONTINUOUS EPITOPES BY PHAGE-DISPLAYED PEPTIDES .2. SELECTION OF CLONES RECOGNIZED BY A PROTECTIVE MONOCLONAL-ANTIBODY AGAINST THE BORDETELLA-PERTUSSIS TOXIN FROM PHAGE PEPTIDE LIBRARIES [J].
FELICI, F ;
LUZZAGO, A ;
FOLGORI, A ;
CORTESE, R .
GENE, 1993, 128 (01) :21-27
[10]   Shear-dependent rolling on von Willebrand factor of mammalian cells expressing the platelet glycoprotein Ib-IX-V complex [J].
Fredrickson, BJ ;
Dong, JF ;
McIntire, LV ;
López, JA .
BLOOD, 1998, 92 (10) :3684-3693