Structure and binding determinants of the recombinant kringle-2 domain of human plasminogen to an internal peptide from a group A Streptococcal surface protein

被引:56
作者
Rios-Steiner, JL
Schenone, M
Mochalkin, I
Tulinsky, A
Castellino, FJ [1 ]
机构
[1] Univ Notre Dame, Dept Chem & Biochem, Notre Dame, IN 46556 USA
[2] Michigan State Univ, Dept Chem, E Lansing, MI 48824 USA
关键词
plasminogen kringle-2; X-ray crystal structure; kringle domains; lysine-binding sites; ligand-binding;
D O I
10.1006/jmbi.2001.4646
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 X-ray crystal structure of a complex of a modified recombinant kringle-2 domain of human plasminogen, K2(Pg)[C(4)G/(ED)-D-56/L(72)y] (MK2(Pg)), containing an upregulated lysine-binding site, bound to a functional 30 residue internal peptide (VEK-30) from an M-type protein of a group A Streptococcus surface protein, has been determined by molecular replacement methods using K4(Pg) as a model, and refined at 2.7 Angstrom resolution to a X-factor of 19.5 %. The X-ray crystal structure shows that VEK-30 exists as a nearly end-to-end alpha -helix in the complex with mK2(Pg). The final structure also revealed that Arg17 and His18 of VEK-30 served as cationic loci for Asp54 and Asp56 of the consensus lysine-binding site of mK2(Pg), while Glu20 of VEK-30 coordinates with Arg69 of the cationic binding site of mK2(Pg). The hydrophobic ligand-binding pocket in mK2(Pg), consisting primarily of Trp60 and Trp70, situated between the positive and negative centers of the lysine-binding site, is utilized in a novel manner in stabilizing the interaction with VEK-30 by forming a cation-pi -electron-mediated association with the positive side-chain of Arg17 of this peptide. Additional lysine-binding sites, as well as exosite electrostatic and hydrogen bonding interactions involving Glu9 and Lys14 of VEK-30, were observed in the structural model. The importance of these interactions were tested in solution by investigating the-binding constants of synthetic variants of VEK-30 to mK2(Pg), and it was found that, Lys14, Arg17, His18, and Glu20 of VEK-30 were the most critical amino acid binding determinants. With regard to the solution studies, circular dichroism analysis of the titration of VEK-30 with mK2(Pg) demonstrated that the peptidic a-helical structure increased substantially when bound to the kringle module, in agreement with the X-ray results. This investigation is the first to delineate structurally the mode of interaction of the lysine-binding site of a kringle with an internal pseudo-lysine residue of a peptide or protein that functionally interacts with a kringle module, and serves as a paradigm for this important class of interactions. (C) 2001 Academic Press.
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页码:705 / 719
页数:15
相关论文
共 63 条
[41]  
Rost B, 1996, METHOD ENZYMOL, V266, P525
[42]  
ROUSSEL A, 1991, SILICON GRAPHICS COR, V91
[43]   An internal histidine residue from the bacterial surface protein, PAM, mediates its binding to the kringle-2 domain of human plasminogen [J].
Schenone, MM ;
Warder, SE ;
Martin, JA ;
Prorok, M ;
Castellino, FJ .
JOURNAL OF PEPTIDE RESEARCH, 2000, 56 (06) :438-445
[44]   DIRECTIONALITY OF THE CATION-PI EFFECT - A CHARGE-MEDIATED SIZE-SELECTIVITY IN BINDING [J].
SCHWABACHER, AW ;
ZHANG, SH ;
DAVY, W .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1993, 115 (15) :6995-6996
[45]  
SEHL LC, 1990, J BIOL CHEM, V265, P5482
[46]   THE CA2+ ION AND MEMBRANE-BINDING STRUCTURE OF THE GLA DOMAIN OF CA-PROTHROMBIN FRAGMENT-1 [J].
SORIANOGARCIA, M ;
PADMANABHAN, K ;
DEVOS, AM ;
TULINSKY, A .
BIOCHEMISTRY, 1992, 31 (09) :2554-2566
[47]  
Sottrup-Jensen L., 1978, Progress in Chemical Fibrinolysis and Thrombolysis, P191
[48]   BINDING OF HUMAN PLASMINOGEN TO BASEMENT-MEMBRANE (TYPE-IV) COLLAGEN [J].
STACK, MS ;
MOSER, TL ;
PIZZO, SV .
BIOCHEMICAL JOURNAL, 1992, 284 :103-108
[49]   CONCERNING THE THERMODYNAMICS OF MOLECULAR RECOGNITION IN AQUEOUS AND ORGANIC MEDIA - EVIDENCE FOR SIGNIFICANT HEAT-CAPACITY EFFECTS [J].
STAUFFER, DA ;
BARRANS, RE ;
DOUGHERTY, DA .
JOURNAL OF ORGANIC CHEMISTRY, 1990, 55 (09) :2762-2767
[50]   SECONDARY-SITE BINDING OF GLU-PLASMIN, LYS-PLASMIN AND MINIPLASMIN TO FIBRIN [J].
SUENSON, E ;
THORSEN, S .
BIOCHEMICAL JOURNAL, 1981, 197 (03) :619-628