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

J. Biol. Chem., Vol. 277, Issue 26, 23807-23814, June 28, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/26/23807    most recent M202107200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Valentini, G. Articles by Mattevi, A. Search for Related Content PubMed PubMed Citation Articles by Valentini, G. Articles by Mattevi, A. Social Bookmarking                      What's this?

Structure and Function of Human Erythrocyte Pyruvate Kinase
MOLECULAR BASIS OF NONSPHEROCYTIC HEMOLYTIC ANEMIA^*

Giovanna Valentini^§¶, Laurent R. Chiarelli^§, Riccardo Fortin, Manuela Dolzan, Alessandro Galizzi, Donald J. Abraham, Changqing Wang, Paola Bianchi^**, Alberto Zanella^**, and Andrea Mattevi

From the  Dipartimento di Genetica e Microbiologia, Università di Pavia, via Abbiategrasso 207, 27100 Pavia, Italy, the ^§ Dipartimento di Biochimica, Università di Pavia, via Taramelli 3b, 27100 Pavia, Italy, the  Department of Medicinal Chemistry, Virginia Commonwealth University, Richmond, Virginia 23219, and the ^** Divisione di Ematologia, IRCCS Ospedale Maggiore di Milano, via Francesco Sforza 35, 20122 Milano, Italy

Deficiency of human erythrocyte isozyme (RPK) is, together with glucose-6-phosphate dehydrogenase deficiency, the most common cause of the nonspherocytic hemolytic anemia. To provide a molecular framework to the disease, we have solved the 2.7 Å resolution crystal structure of human RPK in complex with fructose 1,6-bisphosphate, the allosteric activator, and phosphoglycolate, a substrate analogue, and we have functionally and structurally characterized eight mutants (G332S, G364D, T384M, D390N, R479H, R486W, R504L, and R532W) found in RPK-deficient patients. The mutations target distinct regions of RPK structure, including domain interfaces and catalytic and allosteric sites. The mutations affect to a different extent thermostability, catalytic efficiency, and regulatory properties. These studies are the first to correlate the clinical symptoms with the molecular properties of the mutant enzymes. Mutations greatly impairing thermostability and/or activity are associated with severe anemia. Some mutant proteins exhibit moderate changes in the kinetic parameters, which are sufficient to cause mild to severe anemia, underlining the crucial role of RPK for erythrocyte metabolism. Prediction of the effects of mutations is difficult because there is no relation between the nature and location of the replaced amino acid and the type of molecular perturbation. Characterization of mutant proteins may serve as a valuable tool to assist with diagnosis and genetic counseling.

The atomic coordinates and the structure factors (code 1LIU, 1LIW, 1LIX, and 1LIY) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Zanella, P. Bianchi, and E. Fermo Pyruvate kinase deficiency Haematologica, June 1, 2007; 92(6): 721 - 723. [Full Text] [PDF]

				R. van Wijk and W. W. van Solinge The energy-less red blood cell is lost: erythrocyte enzyme abnormalities of glycolysis Blood, December 15, 2005; 106(13): 4034 - 4042. [Abstract] [Full Text] [PDF]

				A. Diez, F. Gilsanz, J. Martinez, S. Perez-Benavente, N. W. Meza, and J. M. Bautista Life-threatening nonspherocytic hemolytic anemia in a patient with a null mutation in the PKLR gene and no compensatory PKM gene expression Blood, September 1, 2005; 106(5): 1851 - 1856. [Abstract] [Full Text] [PDF]

				L. R. Chiarelli, P. Bianchi, E. Fermo, A. Galizzi, P. Iadarola, A. Mattevi, A. Zanella, and G. Valentini Functional analysis of pyrimidine 5'-nucleotidase mutants causing nonspherocytic hemolytic anemia Blood, April 15, 2005; 105(8): 3340 - 3345. [Abstract] [Full Text] [PDF]

				J. T. Prchal and X. T. Gregg Red Cell Enzymes Hematology, January 1, 2005; 2005(1): 19 - 23. [Abstract] [Full Text] [PDF]