NMR-STUDIES OF THE DYNAMICS OF THE MULTIDOMAIN PROTEIN UROKINASE-TYPE PLASMINOGEN-ACTIVATOR

被引:25
作者
NOWAK, UK
LI, X
TEUTEN, AJ
SMITH, RAG
DOBSON, CM
机构
[1] UNIV OXFORD,OXFORD CTR MOLEC SCI,S PARKS RD,OXFORD OX1 3QR,ENGLAND
[2] UNIV OXFORD,INORGAN CHEM LAB,OXFORD OX1 3QR,ENGLAND
[3] SMITHKLINE BEECHAM PHARMACEUT,EPSOM KT18 5XQ,SURREY,ENGLAND
关键词
D O I
10.1021/bi00052a038
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
u-PA (urokinase-type plasminogen activator or urokinase) has been studied under a variety of solution conditions by 1-D and 2-D NMR spectroscopy. Very high quality spectra could be obtained from the recombinant protein despite the high molecular mass (46 kDa) by appropriate choice of solution conditions; mildly acidic pH and low ionic strength were found to be optimal. Comparison of spectra of u-PA with spectra of the isolated kringle and protease domains, the EGF-kringle pair, and a synthetic peptide from the kringle-protease linker region, enabled sequential assignments in the u-PA spectrum to be made for kringle resonances, and domain-specific assignments for many others. Simulations of line shapes in both 1-D and 2-D spectra enabled effective correlation times for the different domains, both isolated and in the intact protein, to be determined. These have permitted a model of the u-PA dynamics to be put forward involving extensive, but not unrestricted, motion between the different domains.
引用
收藏
页码:298 / 309
页数:12
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