JBC Origene Your Gene Company

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M101550200 on March 12, 2001

J. Biol. Chem., Vol. 276, Issue 21, 18249-18256, May 25, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/21/18249    most recent
M101550200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ikushiro, H.
Right arrow Articles by Kagamiyama, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ikushiro, H.
Right arrow Articles by Kagamiyama, H.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

A Water-soluble Homodimeric Serine Palmitoyltransferase from Sphingomonas paucimobilis EY2395T Strain
PURIFICATION, CHARACTERIZATION, CLONING, AND OVERPRODUCTION*

Hiroko Ikushiro, Hideyuki Hayashi, and Hiroyuki KagamiyamaDagger

From the Department of Biochemistry, Osaka Medical College, Takatsuki, Osaka 569-8686, Japan

Serine palmitoyltransferase (SPT, EC 2.3.1.50) is a key enzyme in sphingolipid biosynthesis and catalyzes the decarboxylative condensation of L-serine and palmitoyl-coenzyme A to 3-ketodihydrosphingosine. We found that the Gram-negative obligatory aerobic bacteria Sphingomonas paucimobilis EY2395T have significant SPT activity and purified SPT to homogeneity. This enzyme is a water-soluble homodimeric protein unlike eukaryotic enzymes, known as heterodimers composed of tightly membrane-bound subunits, named LCB1 and LCB2. The purified SPT shows an absorption spectrum characteristic of a pyridoxal 5'-phosphate-dependent enzyme. The substrate specificity of the Sphingomonas SPT is less strict than the SPT complex from Chinese hamster ovary cells. We isolated the SPT gene encoding 420 amino acid residues (Mr 45,041) and succeeded in overproducing the SPT protein in Escherichia coli, in which the product amounted to about 10-20% of the total protein of the cell extract. Sphingomonas SPT shows about 30% homology with the enzymes of the alpha -oxamine synthase family, and amino acid residues supposed to be involved in catalysis are conserved. The recombinant SPT was catalytically and spectrophotometrically indistinguishable from the native enzyme. This is the first successful overproduction of an active enzyme in the sphingolipid biosynthetic pathway. Sphingomonas SPT is a prototype of the eukaryotic enzyme and would be a useful model to elucidate the reaction mechanism of SPT.

* This work was supported by a Research Grant from the Japan Society for the Promotion of Science ("Research for the Future" Program).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed. Tel.: 81-726-83-1221 (ext. 2645); Fax: 81-726-84-6516; E-mail: med001@art.osaka-med.ac.jp.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
H. Ikushiro, S. Fujii, Y. Shiraiwa, and H. Hayashi
Acceleration of the Substrate C{alpha} Deprotonation by an Analogue of the Second Substrate Palmitoyl-CoA in Serine Palmitoyltransferase
J. Biol. Chem., March 21, 2008; 283(12): 7542 - 7553.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
H. Ikushiro, M. M. Islam, H. Tojo, and H. Hayashi
Molecular Characterization of Membrane-Associated Soluble Serine Palmitoyltransferases from Sphingobacterium multivorum and Bdellovibrio stolpii
J. Bacteriol., August 1, 2007; 189(15): 5749 - 5761.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Han, K. Gable, L. Yan, M. J. Allen, W. H. Wilson, P. Moitra, J. M. Harmon, and T. M. Dunn
Expression of a Novel Marine Viral Single-chain Serine Palmitoyltransferase and Construction of Yeast and Mammalian Single-chain Chimera
J. Biol. Chem., December 29, 2006; 281(52): 39935 - 39942.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Hornemann, S. Richard, M. F. Rutti, Y. Wei, and A. von Eckardstein
Cloning and Initial Characterization of a New Subunit for Mammalian Serine-palmitoyltransferase
J. Biol. Chem., December 8, 2006; 281(49): 37275 - 37281.
[Abstract] [Full Text] [PDF]

Home page
 Plant CellHome page
M. Chen, G. Han, C. R. Dietrich, T. M. Dunn, and E. B. Cahoon
The Essential Nature of Sphingolipids in Plants as Revealed by the Functional Identification and Characterization of the Arabidopsis LCB1 Subunit of Serine Palmitoyltransferase
PLANT CELL, December 1, 2006; 18(12): 3576 - 3593.
[Abstract] [Full Text] [PDF]

Home page
 Protein Sci.Home page
J. Zhang, A. V. Cheltsov, and G. C. Ferreira
Conversion of 5-aminolevulinate synthase into a more active enzyme by linking the two subunits: Spectroscopic and kinetic properties
Protein Sci., May 1, 2005; 14(5): 1190 - 1200.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Yasuda, M. Nishijima, and K. Hanada
Localization, Topology, and Function of the LCB1 Subunit of Serine Palmitoyltransferase in Mammalian Cells
J. Biol. Chem., January 31, 2003; 278(6): 4176 - 4183.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Zhang and G. C. Ferreira
Transient State Kinetic Investigation of 5-Aminolevulinate Synthase Reaction Mechanism
J. Biol. Chem., November 15, 2002; 277(47): 44660 - 44669.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
A. Keck, J. Rau, T. Reemtsma, R. Mattes, A. Stolz, and J. Klein
Identification of Quinoide Redox Mediators That Are Formed during the Degradation of Naphthalene-2-Sulfonate by Sphingomonas xenophaga BN6
Appl. Envir. Microbiol., September 1, 2002; 68(9): 4341 - 4349.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. H. Merrill Jr.
De Novo Sphingolipid Biosynthesis: A Necessary, but Dangerous, Pathway
J. Biol. Chem., July 12, 2002; 277(29): 25843 - 25846.
[Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Gable, G. Han, E. Monaghan, D. Bacikova, M. Natarajan, R. Williams, and T. M. Dunn
Mutations in the Yeast LCB1 and LCB2 Genes, Including Those Corresponding to the Hereditary Sensory Neuropathy Type I Mutations, Dominantly Inactivate Serine Palmitoyltransferase
J. Biol. Chem., March 15, 2002; 277(12): 10194 - 10200.
[Abstract] [Full Text] [PDF]

Home page
 Plant Cell PhysiolHome page
K. Tamura, N. Mitsuhashi, I. Hara-Nishimura, and H. Imai
Characterization of an Arabidopsis cDNA Encoding a Subunit of Serine Palmitoyltransferase, the Initial Enzyme in Sphingolipid Biosynthesis
Plant Cell Physiol., November 1, 2001; 42(11): 1274 - 1281.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.