title: Dynamics and functional differences between dendroaspin and rhodostomin: insights into protein scaffolds in integrin recognition.
abstract: 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 nM. 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 IC₅₀ values of 239.8 and 256.8 nM. 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.
authors.affiliation: Department of Biochemistry and Institute of Basic Medical Sciences, National Cheng Kung University College of Medicine, Tainan 701, Taiwan.
authors.forename: Chun-Ho
authors.initials: CH
authors.lastname: Cheng
authors.forename: Yi-Chun
authors.initials: YC
authors.lastname: Chen
authors.forename: Jia-Hau
authors.initials: JH
authors.lastname: Shiu
authors.forename: Yao-Tsung
authors.initials: YT
authors.lastname: Chang
authors.forename: Yung-Sheng
authors.initials: YS
authors.lastname: Chang
authors.forename: Chun-Hau
authors.initials: CH
authors.lastname: Huang
authors.forename: Chiu-Yueh
authors.initials: CY
authors.lastname: Chen
authors.forename: Woei-Jer
authors.initials: WJ
authors.lastname: Chuang
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D004546:Elapid Venoms / Q000737:chemistry* / Q000494:pharmacology*; D006801:Humans; D039081:Integrin alpha5beta1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D039302:Integrin alphaVbeta3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D016023:Integrins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000494:pharmacology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000037:antagonists & inhibitors / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D012910:Snake Venoms / Q000737:chemistry* / Q000494:pharmacology*
chemical_list: D004546:Elapid Venoms; D039081:Integrin alpha5beta1; D039302:Integrin alphaVbeta3; D016023:Integrins; D009842:Oligopeptides; D010455:Peptides; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011994:Recombinant Proteins; D012910:Snake Venoms; C072026:rhodostomin; C074772:mambin; C047981:arginyl-glycyl-aspartic acid