1H NMR assignment and secondary structure of recombinant RGD-hirudin.
The conformation of a new recombinant RGD-hirudin, which has the activities of anti-thrombin and anti-platelet aggregation, was investigated by multi-dimensional NMR spectroscopy. The 1H NMR spectra of this protein are assigned in a sequential manner by using a combination of 2D NMR techniques to demonstrate through-bond and through-space (<5 A) connectivities. The secondary structure of recombinant RGD-hirudin was deduced from chemical shift indices, sequential NOEs and 3J(HNalpha) coupling constants. The results show that the recombinant RGD-hirudin has two anti-parallel beta-sheets and no alpha-helix, and also that the Arg-Gly-Asp (RGD) binding motif of this protein is located at the end of a long arm, which consists of two anti-parallel beta-strands (residues 26-31 and 36-41). As the strands are connected by a beta-turn, the recombinant RGD-hirudin acquires high flexibility and inhibits platelet aggregation more effectively.
Liu
Xingang
X
Center of Analysis and Measurement, Fudan University, Shanghai 200433, China.
Yan
Xiaomin
X
Mo
Wei
W
Song
Houyan
H
Dai
Linsen
L
D006629:Hirudins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods* / Q000592:standards*; D009842:Oligopeptides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011522:Protons; D011994:Recombinant Proteins / Q000737:chemistry; D012015:Reference Standards
D006629:Hirudins; D009842:Oligopeptides; D011522:Protons; D011994:Recombinant Proteins