The N-terminal epidermal growth factor-like domain of coagulation factor IX -: Probing its functions in the activation of factor IX and factor X with a monoclonal antibody

被引:15
作者
Persson, KEM
Villoutreix, BO
Thämlitz, AM
Knobe, KE
Stenflo, J [1 ]
机构
[1] Lund Univ, Univ Hosp, Dept Clin Chem, S-20502 Malmo, Sweden
[2] Lund Univ, Univ Hosp, Dept Pediat, S-20502 Malmo, Sweden
[3] Univ Paris 05, INSERM, U428, F-75006 Paris, France
关键词
D O I
10.1074/jbc.M205930200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The absence or reduced activity of coagulation factor IX (FIX) causes the severe bleeding disorder hemophilia B. FIX contains an N-terminal Gla domain followed by two epidermal growth factor-like (EGF) domains and a serine protease domain. In this study, the epitope of monoclonal antibody AW, which is directed against the C-terminal part of the first EGF domain in human FIX, was defined, and the antibody was used to study interactions between the EGF domain of FIX and other coagulation proteins. Antibody AW completely blocks activation of FIX by activated factor XI, but activation by activated factor FVII-tissue factor is inhibited only slightly. The antibody also causes a marginal reduction in the apparent k(cat) for factor X both in the presence and absence of activated factor VIII. Based on these results, we produced a preliminary model of the structure of the activated factor IX-activated factor VIII-AW complex on the surface of phospholipid. The model suggests that in the Xase complex, EGF1 of activated factor IX is not involved in direct binding to activated factor VIII. Studies of the interaction of antibody AW with a mutated FIX molecule (R94D) also suggest that the Glu(78)-Arg(94) salt bridge is not important for maintaining the structure of FIX.
引用
收藏
页码:35616 / 35624
页数:9
相关论文
共 44 条
[11]   Vitamin K-dependent biosynthesis of γ-carboxyglutamic acid [J].
Furie, B ;
Bouchard, BA ;
Furie, BC .
BLOOD, 1999, 93 (06) :1798-1808
[12]   Haemophilia B: database of point mutations and short additions and deletions - eighth edition [J].
Giannelli, F ;
Green, PM ;
Sommer, SS ;
Poon, MC ;
Ludwig, M ;
Schwaab, A ;
Reitsma, PH ;
Goossens, M ;
Yoshioka, A ;
Figueiredo, MS ;
Brownlee, GG .
NUCLEIC ACIDS RESEARCH, 1998, 26 (01) :265-268
[13]  
HAMAGUCHI N, 1994, THROMB HAEMOSTASIS, V72, P856
[14]   KEY RESIDUES INVOLVED IN CALCIUM-BINDING MOTIFS IN EGF-LIKE DOMAINS [J].
HANDFORD, PA ;
MAYHEW, M ;
BARON, M ;
WINSHIP, PR ;
CAMPBELL, ID ;
BROWNLEE, GG .
NATURE, 1991, 351 (6322) :164-167
[15]   An Arg Ser substitution in the second epidermal growth factor-like module of factor IX introduces an O-linked carbohydrate and markedly impairs activation by factor XIa and factor VIIa tissue factor and catalytic efficiency of factor IXa [J].
Hertzberg, MS ;
Facey, SL ;
Hogg, PJ .
BLOOD, 1999, 94 (01) :156-163
[16]   Coagulation factor IXa: the relaxed conformation of Tyr99 blocks substrate binding [J].
Hopfner, KP ;
Lang, A ;
Karcher, A ;
Sichler, K ;
Kopetzki, E ;
Brandstetter, H ;
Huber, R ;
Bode, W ;
Engh, RA .
STRUCTURE, 1999, 7 (08) :989-996
[17]   The mechanism of an inhibitory antibody on TF-initiated blood coagulation revealed by the crystal structures of human tissue factor, Fab5G9 and TF•5G9 complex [J].
Huang, MD ;
Syed, R ;
Stura, EA ;
Stone, MJ ;
Stefanko, RS ;
Ruf, W ;
Edgington, TS ;
Wilson, IA .
JOURNAL OF MOLECULAR BIOLOGY, 1998, 275 (05) :873-894
[18]   Structural basis for chemical inhibition of human blood coagulation factor Xa [J].
Kamata, K ;
Kawamoto, H ;
Honma, T ;
Iwama, T ;
Kim, SH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (12) :6630-6635
[19]   Regions 301-303 and 333-339 in the catalytic domain of blood coagulation Factor IX are Factor VIII-interactive sites involved in stimulation of enzyme activity [J].
Kolkman, JA ;
Lenting, PJ ;
Mertens, K .
BIOCHEMICAL JOURNAL, 1999, 339 :217-221
[20]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+