The NMR solution structure of recombinant RGD-hirudin.
The solution structure of a new recombinant RGD-hirudin, which has the activities of anti-thrombin and anti-platelet aggregation, was determined by (1)H nuclear magnetic resonance spectroscopy and compared with the conformations of recombinant wild-type hirudin and hirudin (variant 2, Lys47) of the hirudin thrombin complex. On the basis of total 1284 distance and dihedral angle constraints derived from a series of NMR spectra, 20 conformers were computed with ARIA/CNS programs. The structure of residues 3-30 and 37-48 form a molecular core with two antiparallel beta-sheets as the other two hirudins. However, significant differences were found in the surface electrostatic charge distributions among the three hirudins, especially in the RGD segment of recombinant RGD-hirudin. This difference may be greatly beneficial to its additional function of anti-platelet aggregation. The difference in extended C-terminal makes its both ionic and hydrophobic interactions with the fibrinogen recognition exosite of thrombin more effective.
Song
Xia
X
Center of Analysis and Measurement, Fudan University, Shanghai 200433, China.
Mo
Wei
W
Liu
Xingang
X
Zhu
Lina
L
Yan
Xiaomin
X
Song
Houyan
H
Dai
Linsen
L
D003198:Computer Simulation; D006629:Hirudins / Q000737:chemistry* / Q000648:ultrastructure*; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D010446:Peptide Fragments / Q000737:chemistry* / Q000648:ultrastructure*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000648:ultrastructure
D006629:Hirudins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C069897:hirudin (53-64), RGD-