Neutron diffraction studies on rubredoxin from Pyrococcus furiosus

被引:6
作者
Bau, R [1 ]
机构
[1] Univ So Calif, Dept Chem, Los Angeles, CA 90007 USA
关键词
neutron diffraction; rubredoxin; pyrococcus furiosus; hydrogen bonds; hydrogen-deuterium exchange;
D O I
10.1107/S0909049503024178
中图分类号
TH7 [仪器、仪表];
学科分类号
0804 ; 080401 ; 081102 ;
摘要
Single-crystal neutron diffraction data up to a resolution of 1.5 Angstrom have been collected at room temperature on two forms of rubredoxin using the BIX-3 diffractometer at the JRR-3 reactor of the Japan Atomic Energy Research Institute (JAERI). Rubredoxin is a small iron-sulfur redox protein with 53 amino acid residues, and the source of this particular protein is the hyperthermophile Pyrococcus furiosus, a microorganism that normally lives at temperatures near that of boiling water. Data were collected on crystals of the wild-type protein and on a mutant in which three of the residues have been replaced. In this paper we will be describing several sets of results arising from these high-resolution neutron structure determinations: (a) the H/D exchange pattern of the N-H bonds of the main backbone, which give information about which regions of the molecule are more exposed to solvent, (b) the orientations of some of the O-D bonds in the protein, information which is often not obtainable from X-ray results; (c) the structure and appearance of water molecules in the protein crystals; and (d) some structural features which may help rationalize the remarkable thermal stability of the wild-type protein from this intriguing microorganism.
引用
收藏
页码:76 / 79
页数:4
相关论文
共 17 条
[1]   Crystal structure of rubredoxin from Pyrococcus furiosus at 0.95 Å resolution, and the structures of N-terminal methionine and formylmethionine variants of Pf Rd.: Contributions of N-terminal interactions to thermostability [J].
Bau, R ;
Rees, DC ;
Kurtz, DM ;
Scott, RA ;
Huang, HS ;
Adams, MWW ;
Eidsness, MK .
JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY, 1998, 3 (05) :484-493
[2]  
BRUNGER AT, 1992, X PLOR VERSION 3 1 S
[3]  
CHATAKE T, 2003, UNPUB J AM CHEM SOC
[4]  
CHATAKE T, 2003, IN PRESS J SYNCHROTR
[5]   X-RAY CRYSTAL-STRUCTURES OF THE OXIDIZED AND REDUCED FORMS OF THE RUBREDOXIN FROM THE MARINE HYPERTHERMOPHILIC ARCHAEBACTERIUM PYROCOCCUS-FURIOSUS [J].
DAY, MW ;
HSU, BT ;
JOSHUATOR, L ;
PARK, JB ;
ZHOU, ZH ;
ADAMS, MWW ;
REES, DC .
PROTEIN SCIENCE, 1992, 1 (11) :1494-1507
[6]   Millisecond time scale conformational flexibility in a hyperthermophile protein at ambient temperature [J].
Hernández, G ;
Jenney, FE ;
Adams, MWW ;
LeMaster, DM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (07) :3166-3170
[7]   Stability and dynamics in a hyperthermophilic protein with melting temperature close to 200 degrees C [J].
Hiller, R ;
Zhou, ZH ;
Adams, MWW ;
Englander, SW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (21) :11329-11332
[8]  
Jenney FE, 2001, METHOD ENZYMOL, V334, P45
[9]  
KURIHARA K, 2003, UNPUB
[10]  
KURIHARA K, 2001, J PHYS SOC JPN, V70, P400