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

Nature Structural Biology volume 5, pages 280–283 (1998)Cite this article

Abstract

Here we present a time-resolved crystallographic analysis of the hydrolysis of exo (Sp) guanosine 2′,3′-cyclophosphoroth-ioate 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.

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Authors and Affiliations

  1. Laboratorium voor Ultrastructuur, Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit Brussel, Paardenstraat 65, B-1640 St., Genesius Rode, Belgium

    Ingrid Zegers, Remy Loris, Gillis Dehollander, Abdel Fattah Haikal, Jan Steyaert & Lode Wyns

  2. Vlaamse Instelling voor Technologisch Onderzoek - VITO, Boeretang 200, B-2400, Mol, Belgium

    Freddy Poortmans

Authors
  1. Ingrid Zegers
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  2. Remy Loris
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  3. Gillis Dehollander
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  4. Abdel Fattah Haikal
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  5. Freddy Poortmans
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  6. Jan Steyaert
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  7. Lode Wyns
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Correspondence to Ingrid Zegers.

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Zegers, I., Loris, R., Dehollander, G. et al. Hydrolysis of a slow cyclic thiophosphate substrate of RNase T1 analyzed by time-resolved crystallograph. Nat Struct Mol Biol 5, 280–283 (1998). https://doi.org/10.1038/nsb0498-280

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