HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 33, 34691-34697, August 13, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/33/34691    most recent M405400200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bjelke, J. R. Articles by Rasmussen, H. B. Search for Related Content PubMed PubMed Citation Articles by Bjelke, J. R. Articles by Rasmussen, H. B. Social Bookmarking                      What's this?

Tyrosine 547 Constitutes an Essential Part of the Catalytic Mechanism of Dipeptidyl Peptidase IV^*

Jais R. Bjelke, Jesper Christensen, Sven Branner¶, Nicolai Wagtmann||, Christina Olsen||, Anders B. Kanstrup**, and Hanne B. Rasmussen

From the Institute of Medical Biochemistry and Genetics, Panum Institute, University of Copenhagen, Bledamsvej 3C, DK-2200 Copenhagen and ^¶Protein Science, ^||Cancer and Immunobiology, ^**Medicinal Chemistry, and Protein Structure, Novo Nordisk A/S, Novo Allé, DK-2880 Bagsvaerd, Denmark

Human dipeptidyl peptidase IV (DPP-IV) is a ubiquitously expressed type II transmembrane serine protease. It cleaves the penultimate positioned prolyl bonds at the N terminus of physiologically important peptides such as the incretin hormones glucagon-like peptide 1 and glucose-dependent insulinotropic peptide. In this study, we have characterized different active site mutants. The Y547F mutant as well as the catalytic triad mutants S630A, D708A, and H740L showed less than 1% wild type activity. X-ray crystal structure analysis of the Y547F mutant revealed no overall changes compared with wild type apoDPP-IV, except the ablation of the hydroxyl group of Tyr⁵⁴⁷ and a water molecule positioned in close proximity to Tyr⁵⁴⁷. To elucidate further the reaction mechanism, we determined the crystal structure of DPP-IV in complex with diisopropyl fluorophosphate, mimicking the tetrahedral intermediate. The kinetic and structural findings of the tyrosine residue are discussed in relation to the catalytic mechanism of DPP-IV and to the inhibitory mechanism of the 2-cyanopyrrolidine class of potent DPP-IV inhibitors, proposing an explanation for the specificity of this class of inhibitors for the S9b family among serine proteases.

Received for publication, May 14, 2004 , and in revised form, June 1, 2004.

The atomic coordinates and structure factors (codes ITO7, ITK3, and ITKR) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Present address: Laboratory for Molecular Pharmacology, Panum Institute, University of Copenhagen, Blegdamsvej 3C, DK-2200 Copenhagen N, Denmark.

To whom correspondence should be addressed. E-mail: hbrm{at}novonordisk.com.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				W. J. Metzler, J. Yanchunas, C. Weigelt, K. Kish, H. E. Klei, D. Xie, Y. Zhang, M. Corbett, J. K. Tamura, B. He, et al. Involvement of DPP-IV catalytic residues in enzyme-saxagliptin complex formation Protein Sci., February 1, 2008; 17(2): 240 - 250. [Abstract] [Full Text] [PDF]

				D. Chen, A. Kennedy, J.-Y. Wang, W. Zeng, Q. Zhao, M. Pearl, M. Zhang, Z. Suo, J. M. Nesland, Y. Qiao, et al. Activation of EDTA-Resistant Gelatinases in Malignant Human Tumors. Cancer Res., October 15, 2006; 66(20): 9977 - 9985. [Abstract] [Full Text] [PDF]