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Dynamics and functional differences between dendroaspin and rhodostomin: Insights into protein scaffolds in integrin recognition
Author(s) -
Cheng ChunHo,
Chen YiChun,
Shiu JiaHau,
Chang YaoTsung,
Chang YungSheng,
Huang ChunHau,
Chen ChiuYueh,
Chuang WoeiJer
Publication year - 2012
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.2169
Subject(s) - integrin , rgd motif , recombinant dna , disintegrin , chemistry , pichia pastoris , biophysics , docking (animal) , microbiology and biotechnology , biochemistry , biology , cell , matrix metalloproteinase , gene , medicine , metalloproteinase , nursing
Dendroaspin (Den) and rhodostomin (Rho) are snake venom proteins containing a PRGDMP motif. Although Den and Rho have different 3D structures, they are highly potent integrin inhibitors . To study their structure, function, and dynamics relationships, we expressed Den and Rho in Pichia pastoris . The recombinant Den and Rho inhibited platelet aggregation with the K I values of 149.8 and 83.2 n M . Cell adhesion analysis showed that Den was 3.7 times less active than Rho when inhibiting the integrin αIIbβ3 and 2.5 times less active when inhibiting the integrin αvβ3. In contrast, Den and Rho were similarly active when inhibiting the integrin α5β1 with the IC 50 values of 239.8 and 256.8 n M . NMR analysis showed that recombinant Den and Rho have different 3D conformations for their arginyl‐glycyl‐aspartic acid (RGD) motif. However, the comparison with Rho showed that the docking of Den into integrin αvβ3 resulted in a similar number of contacts. Analysis of the dynamic properties of the RGD loop in Den and Rho showed that they also had different dynamic properties. These results demonstrate that protein scaffolds affect the function, structure, and dynamics of their RGD motif.