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

J. Biol. Chem., Vol. 277, Issue 44, 41410-41416, November 1, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/44/41410    most recent M206863200v1 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 Fischer, M. Articles by Bacher, A. Search for Related Content PubMed PubMed Citation Articles by Fischer, M. Articles by Bacher, A. Social Bookmarking                      What's this?

Biosynthesis of Riboflavin in Archaea Studies on the Mechanism of 3,4-Dihydroxy-2-butanone-4-phosphate Synthase of Methanococcus jannaschii^*

Markus Fischer^§, Werner Römisch, Susanne Schiffmann, Mark Kelly^¶, Hartmut Oschkinat^¶, Stefan Steinbacher, Robert Huber, Wolfgang Eisenreich, Gerald Richter, and Adelbert Bacher

From the  Institut für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany,  Department of Protein Crystallography, Max-Planck-Institute of Biochemistry, Am Klopferspitz 18a, D-82512 Martinsried, Germany, and ^¶ Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, D-13125 Berlin, Germany

The hypothetical protein predicted by the open reading frame MJ0055 of Methanococcus jannaschii was expressed in a recombinant Escherichia coli strain under the control of a synthetic gene optimized for translation in an eubacterial host. The recombinant protein catalyzes the formation of the riboflavin precursor 3,4-dihydroxy-2-butanone 4-phosphate from ribulose 5-phosphate at a rate of 174 nmol mg¹ min¹ at 37 °C. The homodimeric 51.6-kDa protein requires divalent metal ions, preferentially magnesium, for activity. The reaction involves an intramolecular skeletal rearrangement as shown by ¹³C NMR spectroscopy using [U-¹³C₅]ribulose 5-phosphate as substrate. A cluster of charged amino acid residues comprising arginine 25, glutamates 26 and 28, and aspartates 21 and 30 is essential for catalytic activity. Histidine 164 and glutamate 185 were also shown to be essential for catalytic activity.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF490541-AF490573 and AF516684.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				M. Fischer, W. Romisch, S. Saller, B. Illarionov, G. Richter, F. Rohdich, W. Eisenreich, and A. Bacher Evolution of Vitamin B2 Biosynthesis: STRUCTURAL AND FUNCTIONAL SIMILARITY BETWEEN PYRIMIDINE DEAMINASES OF EUBACTERIAL AND PLANT ORIGIN J. Biol. Chem., August 27, 2004; 279(35): 36299 - 36308. [Abstract] [Full Text] [PDF]

				S. Steinbacher, S. Schiffmann, G. Richter, R. Huber, A. Bacher, and M. Fischer Structure of 3,4-Dihydroxy-2-butanone 4-Phosphate Synthase from Methanococcus jannaschii in Complex with Divalent Metal Ions and the Substrate Ribulose 5-Phosphate: IMPLICATIONS FOR THE CATALYTIC MECHANISM J. Biol. Chem., October 24, 2003; 278(43): 42256 - 42265. [Abstract] [Full Text] [PDF]