Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis

Ana K. Pedersen, Xiaoling Guo, Karin B. Møller, Günther H. Peters, Henrik S. Andersen, Jette S. Kastrup, Steen B. Mortensen, Lars F. Iversen, Zhong Yin Zhang, Niels Peter H Møller

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Previous enzyme kinetic and structural studies have revealed a critical role for Asp181 (PTP1B numbering) in PTP (protein-tyrosine phosphatase)-mediated catalysis. In the E-P (phosphoenzyme) formation step, Asp181 functions as a general acid, while in the E-P hydrolysis step it acts as a general base. Most of our understanding of the role of Asp 181 is derived from studies with the Yersinia PTP and the mammalian PTP1B, and to some extent also TC (T-cell)-PTP and the related PTPα and PTPε. The neighbouring residue 182 is a phenylalanine in these four mammalian enzymes and a glutamine in Yersinia PTP. Surprisingly, little attention has been paid to the fact that this residue is a histidine in most other mammalian PTPs. Using a reciprocal single-point mutational approach with introduction of His182 in PTP1B and Phe182 in PTPH1, we demonstrate here that His182-PTPs, in comparison with Phe 182-PTPs, have significantly decreased kcat values, and to a lesser degree, decreased kcat/Km values. Combined enzyme kinetic, X-ray crystallographic and molecular dynamics studies indicate that the effect of His182 is due to interactions with Asp 181 and with Gln262. We conclude that residue 182 can modulate the functionality of both Asp181 and Gln262 and therefore affect the E-P hydrolysis step of PTP-mediated catalysis.

Original languageEnglish (US)
Pages (from-to)421-433
Number of pages13
JournalBiochemical Journal
Volume378
Issue number2
DOIs
StatePublished - Mar 1 2004

Fingerprint

Protein Tyrosine Phosphatases
Catalysis
Yersinia
Enzyme kinetics
Non-Receptor Type 2 Protein Tyrosine Phosphatase
Hydrolysis
Enzymes
Molecular Dynamics Simulation
Glutamine
Phenylalanine
Histidine
Molecular dynamics
X-Rays
X rays
Acids

Keywords

  • Enzyme kinetics
  • Molecular dynamics
  • Protein-tyrosine phosphatase
  • PTP1B
  • PTPH1
  • X-ray crystallography

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pedersen, A. K., Guo, X., Møller, K. B., Peters, G. H., Andersen, H. S., Kastrup, J. S., ... Møller, N. P. H. (2004). Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis. Biochemical Journal, 378(2), 421-433. https://doi.org/10.1042/BJ20030565

Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis. / Pedersen, Ana K.; Guo, Xiaoling; Møller, Karin B.; Peters, Günther H.; Andersen, Henrik S.; Kastrup, Jette S.; Mortensen, Steen B.; Iversen, Lars F.; Zhang, Zhong Yin; Møller, Niels Peter H.

In: Biochemical Journal, Vol. 378, No. 2, 01.03.2004, p. 421-433.

Research output: Contribution to journalArticle

Pedersen, AK, Guo, X, Møller, KB, Peters, GH, Andersen, HS, Kastrup, JS, Mortensen, SB, Iversen, LF, Zhang, ZY & Møller, NPH 2004, 'Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis', Biochemical Journal, vol. 378, no. 2, pp. 421-433. https://doi.org/10.1042/BJ20030565
Pedersen AK, Guo X, Møller KB, Peters GH, Andersen HS, Kastrup JS et al. Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis. Biochemical Journal. 2004 Mar 1;378(2):421-433. https://doi.org/10.1042/BJ20030565
Pedersen, Ana K. ; Guo, Xiaoling ; Møller, Karin B. ; Peters, Günther H. ; Andersen, Henrik S. ; Kastrup, Jette S. ; Mortensen, Steen B. ; Iversen, Lars F. ; Zhang, Zhong Yin ; Møller, Niels Peter H. / Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis. In: Biochemical Journal. 2004 ; Vol. 378, No. 2. pp. 421-433.
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abstract = "Previous enzyme kinetic and structural studies have revealed a critical role for Asp181 (PTP1B numbering) in PTP (protein-tyrosine phosphatase)-mediated catalysis. In the E-P (phosphoenzyme) formation step, Asp181 functions as a general acid, while in the E-P hydrolysis step it acts as a general base. Most of our understanding of the role of Asp 181 is derived from studies with the Yersinia PTP and the mammalian PTP1B, and to some extent also TC (T-cell)-PTP and the related PTPα and PTPε. The neighbouring residue 182 is a phenylalanine in these four mammalian enzymes and a glutamine in Yersinia PTP. Surprisingly, little attention has been paid to the fact that this residue is a histidine in most other mammalian PTPs. Using a reciprocal single-point mutational approach with introduction of His182 in PTP1B and Phe182 in PTPH1, we demonstrate here that His182-PTPs, in comparison with Phe 182-PTPs, have significantly decreased kcat values, and to a lesser degree, decreased kcat/Km values. Combined enzyme kinetic, X-ray crystallographic and molecular dynamics studies indicate that the effect of His182 is due to interactions with Asp 181 and with Gln262. We conclude that residue 182 can modulate the functionality of both Asp181 and Gln262 and therefore affect the E-P hydrolysis step of PTP-mediated catalysis.",
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AU - Møller, Karin B.

AU - Peters, Günther H.

AU - Andersen, Henrik S.

AU - Kastrup, Jette S.

AU - Mortensen, Steen B.

AU - Iversen, Lars F.

AU - Zhang, Zhong Yin

AU - Møller, Niels Peter H

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