Impaired transition state complementarity in the hydrolysis of O- arylphosphorothioates by protein-tyrosine phosphatases

Yan Ling Zhang, Florian Hollfelder, Steven J. Gordon, Li Chen, Yen Fang Keng, Li Wu, Daniel Herschlag, Zhong-Yin Zhang

Research output: Contribution to journalArticle

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Abstract

The hydrolysis of O-arylphosphorothioates by protein-tyrosine phosphatases (PTPases) was studied with the aim of providing a mechanistic framework for the reactions of this important class of substrate analogues. O-Arylphosphorothioates are hydrolyzed 2 to 3 orders of magnitude slower than O-aryl phosphates by PTPases. This is in contrast to the solution reaction where phosphorothioates display 10-60-fold higher reactivity than the corresponding oxygen analogues. Kinetic analyses suggest that PTPases utilize the same active site and similar kinetic and chemical mechanisms for the hydrolysis of O-arylphosphorothioates and O-aryl phosphates. Thio substitution has no effect on the affinity of substrate or product for the PTPases. Bronsted analyses suggest that like the PTPase-catalyzed phosphoryl transfer reaction the transition state for the PTPase-catalyzed thiophosphoryl transfer is highly dissociative, similar to that of the corresponding solution reaction. The side chain of the active-site Arg residue forms a bidentate hydrogen bond with two of the terminal phosphate oxygens in the ground state and two of the equatorial oxygens in a transition state analog complex with vanadate [Denu et al. (1996) Proc. Natl. Acad. Sci. USA 93, 2493-2498; Zhang, M. et al. (1997) Biochemistry 36, 15-23; Pannifer et al. (1998) J. Biol. Chem. 273, 10454-10462]. Replacement of the active- site Arg409 in the Yersinia PTPase by a Lys reduces the thio effect by 54- fold, consistent with direct interaction and demonstrating strong energetic coupling between Arg409 and the phosphoryl oxygens in the transition state. These results suggest that the large thio effect observed in the PTPase reaction is the result of inability to achieve precise transition state complementarity in the enzyme active site with the larger sulfur substitution.

Original languageEnglish (US)
Pages (from-to)12111-12123
Number of pages13
JournalBiochemistry
Volume38
Issue number37
DOIs
StatePublished - Sep 14 1999
Externally publishedYes

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Protein Tyrosine Phosphatases
Hydrolysis
Catalytic Domain
Oxygen
Phosphates
Substitution reactions
Yersinia
Biochemistry
Kinetics
Vanadates
Substrates
Sulfur
Ground state
Hydrogen
Hydrogen bonds
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Impaired transition state complementarity in the hydrolysis of O- arylphosphorothioates by protein-tyrosine phosphatases. / Zhang, Yan Ling; Hollfelder, Florian; Gordon, Steven J.; Chen, Li; Keng, Yen Fang; Wu, Li; Herschlag, Daniel; Zhang, Zhong-Yin.

In: Biochemistry, Vol. 38, No. 37, 14.09.1999, p. 12111-12123.

Research output: Contribution to journalArticle

Zhang, YL, Hollfelder, F, Gordon, SJ, Chen, L, Keng, YF, Wu, L, Herschlag, D & Zhang, Z-Y 1999, 'Impaired transition state complementarity in the hydrolysis of O- arylphosphorothioates by protein-tyrosine phosphatases', Biochemistry, vol. 38, no. 37, pp. 12111-12123. https://doi.org/10.1021/bi990836i
Zhang, Yan Ling ; Hollfelder, Florian ; Gordon, Steven J. ; Chen, Li ; Keng, Yen Fang ; Wu, Li ; Herschlag, Daniel ; Zhang, Zhong-Yin. / Impaired transition state complementarity in the hydrolysis of O- arylphosphorothioates by protein-tyrosine phosphatases. In: Biochemistry. 1999 ; Vol. 38, No. 37. pp. 12111-12123.
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AU - Wu, Li

AU - Herschlag, Daniel

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