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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aspergillus oryzae / enzymology
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Binding Sites
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Computer Simulation
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Crystallography, X-Ray / methods
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Cyclic GMP / analogs & derivatives*
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Cyclic GMP / chemistry
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Cyclic GMP / metabolism
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Hydrolysis
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Models, Molecular
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Molecular Conformation
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Molecular Sequence Data
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Protein Conformation
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Ribonuclease T1 / chemistry*
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Ribonuclease T1 / metabolism*
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Substrate Specificity
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Thionucleotides / chemistry
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Thionucleotides / metabolism*
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Time Factors
Substances
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Thionucleotides
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guanosine 2',3'-cyclophosphorothioate
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Ribonuclease T1
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Cyclic GMP
Associated data
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PDB/1GSP
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PDB/2GSP
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PDB/3GSP
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PDB/4GSP
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PDB/5GSP
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PDB/6GSP
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PDB/7GSP