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

This Article Full Text Full Text (PDF) 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 Jordan, D. B. Articles by Viitanen, P. V. Search for Related Content PubMed PubMed Citation Articles by Jordan, D. B. Articles by Viitanen, P. V. Social Bookmarking                      What's this?

J Biol Chem, Vol. 274, Issue 31, 22114-22121, July 30, 1999

Plant Riboflavin Biosynthesis
CLONING, CHLOROPLAST LOCALIZATION, EXPRESSION, PURIFICATION, AND PARTIAL CHARACTERIZATION OF SPINACH LUMAZINE SYNTHASE

Douglas B. Jordan, Karen O. Bacot^§, Thomas J. Carlson^¶, Martin Kessel, and Paul V. Viitanen^§

From the  E. I. DuPont de Nemours Agricultural Products, Stine-Haskell Research Center, Newark, Delaware 19714, the ^§ E. I. DuPont de Nemours Life Sciences, Experimental Station, Wilmington, Delaware 19880-0402, ^¶ E. I. DuPont de Nemours Agricultural Products, Experimental Station, Wilmington, Delaware 19880-0402, and the  Laboratory of Structural Biology, National Institutes of Health, Bethesda, Maryland 20892

Lumazine synthase, which catalyzes the penultimate step of riboflavin biosynthesis, has been cloned from three higher plants (spinach, tobacco, and arabidopsis) through functional complementation of an Escherichia coli auxotroph. Whereas the three plant proteins exhibit some structural similarities to known microbial homologs, they uniquely possess N-terminal polypeptide extensions that resemble typical chloroplast transit peptides. In vitro protein import assays with intact chloroplasts and immunolocalization experiments verify that higher plant lumazine synthase is synthesized in the cytosol as a larger molecular weight precursor protein, which is post-translationally imported into chloroplasts where it is proteolytically cleaved to its mature size. The authentic spinach enzyme is estimated to constitute <0.02% of the total chloroplast protein. Recombinant "mature" spinach lumazine synthase is expressed in E. coli at levels exceeding 30% of the total soluble protein and is readily purified to homogeneity using a simple two-step procedure. Apparent V_max and K_m values obtained with the purified plant protein are similar to those reported for microbial lumazine synthases. Electron microscopy and hydrodynamic studies reveal that native plant lumazine synthase is a hollow capsid-like structure comprised of 60 identical 16.5-kDa subunits, resembling its icosahedral counterparts in E. coli and Bacillus subtilis.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				F. J. Sandoval, Y. Zhang, and S. Roje Flavin Nucleotide Metabolism in Plants: MONOFUNCTIONAL ENZYMES SYNTHESIZE FAD IN PLASTIDS J. Biol. Chem., November 7, 2008; 283(45): 30890 - 30900. [Abstract] [Full Text] [PDF]

				T. Ogawa, K. Yoshimura, H. Miyake, K. Ishikawa, D. Ito, N. Tanabe, and S. Shigeoka Molecular Characterization of Organelle-Type Nudix Hydrolases in Arabidopsis Plant Physiology, November 1, 2008; 148(3): 1412 - 1424. [Abstract] [Full Text] [PDF]

				V. Zylberman, S. Klinke, I. Haase, A. Bacher, M. Fischer, and F. A. Goldbaum Evolution of vitamin b2 biosynthesis: 6,7-dimethyl-8-ribityllumazine synthases of Brucella. J. Bacteriol., September 1, 2006; 188(17): 6135 - 6142. [Abstract] [Full Text] [PDF]

				J.-B. Peltier, Y. Cai, Q. Sun, V. Zabrouskov, L. Giacomelli, A. Rudella, A. J. Ytterberg, H. Rutschow, and K. J. van Wijk The Oligomeric Stromal Proteome of Arabidopsis thaliana Chloroplasts Mol. Cell. Proteomics, January 1, 2006; 5(1): 114 - 133. [Abstract] [Full Text] [PDF]

				F. J. Sandoval and S. Roje An FMN Hydrolase Is Fused to a Riboflavin Kinase Homolog in Plants J. Biol. Chem., November 18, 2005; 280(46): 38337 - 38345. [Abstract] [Full Text] [PDF]

				T. Yoshimura-Suzuki, I. Sagami, N. Yokota, H. Kurokawa, and T. Shimizu DOSEc, a Heme-Regulated Phosphodiesterase, Plays an Important Role in the Regulation of the Cyclic AMP Level in Escherichia coli J. Bacteriol., October 1, 2005; 187(19): 6678 - 6682. [Abstract] [Full Text] [PDF]

				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. Xiao, L. Dai, F. Liu, Z. Wang, W. Peng, and D. Xie COS1: An Arabidopsis coronatine insensitive1 Suppressor Essential for Regulation of Jasmonate-Mediated Plant Defense and Senescence PLANT CELL, May 1, 2004; 16(5): 1132 - 1142. [Abstract] [Full Text] [PDF]

				M. S. Fornasari, D. A. Laplagne, N. Frankel, A. A. Cauerhff, F. A. Goldbaum, and J. Echave Sequence Determinants of Quaternary Structure in Lumazine Synthase Mol. Biol. Evol., January 1, 2004; 21(1): 97 - 107. [Abstract] [Full Text] [PDF]