Chloramine T-induced structural and biochemical changes in echistatin.
Echistatin is a member of the disintegrin family of peptides and a potent inhibitor of platelet aggregation and cell adhesion. Echistatin binds to integrin alpha(v)beta3 and alpha(IIb)beta3 receptors with high affinity. Binding is mediated by an RGD-containing loop maintained in an appropriate conformation by disulfide bridges. In this study, we have compared the binding characteristics of echistatin iodinated by either lactoperoxidase or chloramine T method. We show that echistatin labeled by lactoperoxidase method binds to integrin alpha(v)beta3 receptor with high affinity and in a non-dissociable manner very similar to native echistatin. In contrast, chloramine T-labeled echistatin can rapidly dissociate from the receptor. We demonstrate that chloramine T reaction results in the addition of an extra oxygen to the methionine residue adjacent to the RGD motif in echistatin. Modeling studies and molecular dynamic simulation studies show that the extra oxygen atom on the methionine residue can form hydrogen bonds with the glycine and aspartic acid residues of the RGD motif. These structural changes in echistatin help explain the changes in the binding characteristics of the molecule following chloramine T reaction.
Kumar
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CC
Department of Tumor Biology, Schering-Plough Research Institute, Kenilworth, NJ 07033, USA. Chandra.kumar@spcorp.com
Nie
H
H
Armstrong
L
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Zhang
R
R
Vijay-Kumar
S
S
Tsarbopoulos
A
A
D002700:Chloramines / Q000737:chemistry*; D036341:Intercellular Signaling Peptides and Proteins; D007784:Lactoperoxidase / Q000378:metabolism; D013058:Mass Spectrometry; D008715:Methionine / Q000737:chemistry; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D019030:Receptors, Vitronectin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014105:Tosyl Compounds / Q000737:chemistry*
D002700:Chloramines; D036341:Intercellular Signaling Peptides and Proteins; D009842:Oligopeptides; D010455:Peptides; D010975:Platelet Aggregation Inhibitors; D019030:Receptors, Vitronectin; D014105:Tosyl Compounds; C016300:chloramine-T; C057986:echistatin; C047981:arginyl-glycyl-aspartic acid; D008715:Methionine; D007784:Lactoperoxidase