Hydrolysis of a slow cyclic thiophosphate substrate of RNase T1 analyzed by time-resolved crystallography.

TitleHydrolysis of a slow cyclic thiophosphate substrate of RNase T1 analyzed by time-resolved crystallography.
Publication TypeJournal Article
Year of Publication1998
AuthorsZegers, I., R. Loris, G. Dehollander, F. A Haikal, F. Poortmans, J. Steyaert, and L. Wyns
JournalNat Struct Biol
Volume5
Issue4
Pagination280-3
Date Published1998 Apr
ISSN1072-8368
KeywordsAspergillus oryzae, Binding Sites, Computer Simulation, Crystallography, X-Ray, Cyclic GMP, Hydrolysis, Models, Molecular, Molecular Conformation, Molecular Sequence Data, Protein Conformation, Ribonuclease T1, Substrate Specificity, Thionucleotides, Time Factors
Abstract

Here we present a time-resolved crystallographic analysis of the hydrolysis of exo (Sp) guanosine 2',3'-cyclophosphorothioate by RNase T1. The use of a slow substrate and fast crystallization methods made it possible to perform the study with conventional data-collection techniques. The results support the idea that the hydrolysis reaction proceeds through a mechanism that is the inverse of the transesterification reaction. In addition, the structures provide an explanation for the differential behavior of RNase T1 towards exo- and endo-cyclic thiophosphates.
Alternate JournalNat. Struct. Biol.
PubMed ID9546218