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 Asp181 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 PTP∊. 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 Phe182-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 Asp181 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.
Skip Nav Destination
Article navigation
Research Article|
March 01 2004
Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis
Anja K. PEDERSEN;
Anja K. PEDERSEN
*Protein Chemistry, Novo Nordisk, DK-2880 Bagsvaerd, Denmark
†Signal Transduction, Novo Nordisk, DK-2880 Bagsvaerd, Denmark
‡Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, DK-2100 Copenhagen, Denmark
Search for other works by this author on:
Xiao-Ling GUO;
Xiao-Ling GUO
§Department of Molecular Pharmacology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, U.S.A.
Search for other works by this author on:
Karin B. MØLLER;
Karin B. MØLLER
†Signal Transduction, Novo Nordisk, DK-2880 Bagsvaerd, Denmark
Search for other works by this author on:
Günther H. PETERS;
Günther H. PETERS
‖Department of Chemistry, MEMPHYS – Center for Biomembrane Physics, Technical University of Denmark DK-2880 Lyngby, Denmark
Search for other works by this author on:
Henrik S. ANDERSEN;
Henrik S. ANDERSEN
¶MedChem Research I, Novo Nordisk, DK-2760 Måløv, Denmark
Search for other works by this author on:
Jette S. KASTRUP;
Jette S. KASTRUP
‡Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, DK-2100 Copenhagen, Denmark
Search for other works by this author on:
Steen B. MORTENSEN;
Steen B. MORTENSEN
*Protein Chemistry, Novo Nordisk, DK-2880 Bagsvaerd, Denmark
Search for other works by this author on:
Lars F. IVERSEN;
Lars F. IVERSEN
*Protein Chemistry, Novo Nordisk, DK-2880 Bagsvaerd, Denmark
Search for other works by this author on:
Zhong-Yin ZHANG;
Zhong-Yin ZHANG
1
§Department of Molecular Pharmacology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, U.S.A.
Search for other works by this author on:
Niels Peter H. MØLLER
Niels Peter H. MØLLER
1
†Signal Transduction, Novo Nordisk, DK-2880 Bagsvaerd, Denmark
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
April 15 2003
Revision Received:
September 04 2003
Accepted:
October 23 2003
Accepted Manuscript online:
October 23 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2004
2004
Biochem J (2004) 378 (2): 421–433.
Article history
Received:
April 15 2003
Revision Received:
September 04 2003
Accepted:
October 23 2003
Accepted Manuscript online:
October 23 2003
Citation
Anja K. PEDERSEN, Xiao-Ling 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; Residue 182 influences the second step of protein-tyrosine phosphatase-mediated catalysis. Biochem J 1 March 2004; 378 (2): 421–433. doi: https://doi.org/10.1042/bj20030565
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.