1RHL

RIBONUCLEASE T1 COMPLEXED WITH 2'GMP/G23A MUTANT

1RHL の概要

• エントリーDOI: 10.2210/pdb1rhl/pdb
• 分子名称: PROTEIN (RIBONUCLEASE T1), CALCIUM ION, GUANOSINE-2'-MONOPHOSPHATE, ... (4 entities in total)
• 機能のキーワード: endoribonuclease, ribonuclease, endonuclease, hydrolase
• 由来する生物種: Aspergillus oryzae
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 11512.02

構造登録者

Huyghues-Despointes, B.M.P.,Langhorst, U.,Steyaert, J.,Pace, C.N.,Scholtz, J.M. (登録日: 1998-10-09, 公開日: 1998-10-14, 最終更新日: 2024-10-30)

主引用文献

Huyghues-Despointes, B.M.,Langhorst, U.,Steyaert, J.,Pace, C.N.,Scholtz, J.M.
Hydrogen-exchange stabilities of RNase T1 and variants with buried and solvent-exposed Ala --> Gly mutations in the helix.
Biochemistry, 38:16481-16490, 1999

PubMed Abstract: Hydrogen-exchange rates were measured for RNase T1 and three variants with Ala --> Gly substitutions at a solvent-exposed (residue 21) and a buried (residue 23) position in the helix: A21G, G23A, and A21G + G23A. These results were used to measure the stabilities of the proteins. The hydrogen-exchange stabilities (DeltaG(HX)) for the most stable residues in each variant agree with the equilibrium conformational stability measured by urea denaturation (DeltaG(U)), if the effects of D(2)O and proline isomerization are included [Huyghues-Despointes, B. M. P., Scholtz, J. M., and Pace, C. N. (1999) Nat. Struct. Biol. 6, 210-212]. These residues also show similar changes in DeltaG(HX) upon Ala --> Gly mutations (DeltaDeltaG(HX)) as compared to equilibrium measurements (DeltaDeltaG(U)), indicating that the most stable residues are exchanging from the globally unfolded ensemble. Alanine is stabilizing compared to glycine by 1 kcal/mol at a solvent-exposed site 21 as seen by other methods for the RNase T1 protein and peptide helix [Myers, J. K., Pace, C. N., and Scholtz, J. M. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 3833-2837], while it is destabilizing at the buried site 23 by the same amount. For the A21G variant, only local NMR chemical shift perturbations are observed compared to RNase T1. For the G23A variant, large chemical shift changes are seen throughout the sequence, although X-ray crystal structures of the variant and RNase T1 are nearly superimposable. Ala --> Gly mutations in the helix of RNase T1 at both helical positions alter the native-state hydrogen-exchange stabilities of residues throughout the sequence.

PubMed: 10600109
DOI: 10.1021/bi9919450

実験手法

X-RAY DIFFRACTION (1.95 Å)