Abstract:
:Cyclic 2,3-diphosphoglycerate synthetase (cDPGS) catalyzes the synthesis of cyclic 2,3-diphosphoglycerate (cDPG) by formation of an intramolecular phosphoanhydride bond in 2,3-diphosphoglycerate. cDPG is known to be accumulated to high intracellular concentrations (>300 mM) as a putative thermoadapter in some hyperthermophilic methanogens. For the first time, we have purified active cDPGS from a methanogen, the hyperthermophilic archaeon Methanothermus fervidus, sequenced the coding gene, and expressed it in Escherichia coli. cDPGS purification resulted in enzyme preparations containing two isoforms differing in their electrophoretic mobility under denaturing conditions. Since both polypeptides showed the same N-terminal amino acid sequence and Southern analyses indicate the presence of only one gene coding for cDPGS in M. fervidus, the two polypeptides originate from the same gene but differ by a not yet identified modification. The native cDPGS represents a dimer with an apparent molecular mass of 112 kDa and catalyzes the reversible formation of the intramolecular phosphoanhydride bond at the expense of ATP. The enzyme shows a clear preference for the synthetic reaction: the substrate affinity and the Vmax of the synthetic reaction are a factor of 8 to 10 higher than the corresponding values for the reverse reaction. Comparison with the kinetic properties of the electrophoretically homogeneous, apparently unmodified recombinant enzyme from E. coli revealed a twofold-higher Vmax of the enzyme from M. fervidus in the synthesizing direction.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Matussek K,Moritz P,Brunner N,Eckerskorn C,Hensel Rdoi
10.1128/JB.180.22.5997-6004.1998subject
Has Abstractpub_date
1998-11-01 00:00:00pages
5997-6004issue
22eissn
0021-9193issn
1098-5530journal_volume
180pub_type
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journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
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更新日期:1969-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:1999-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.1128/JB.91.5.1977-1985.1966
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1991-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2001-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.155.1.159-168.1983
更新日期:1983-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1997-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.85.2.382-393.1963
更新日期:1963-02-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1968-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1969-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:1987-02-01 00:00:00
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更新日期:2010-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.176.9.2759-2762.1994
更新日期:1994-05-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:2011-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:1983-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2008-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1973-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1975-01-01 00:00:00
abstract::The two-component regulatory system, OmpR-EnvZ, of Escherichia coli K-12 regulates the expression of the major outer membrane porin protein, OmpF. OmpR is a DNA-binding protein which acts as both an activator and a repressor to control ompF transcription. In this article, we describe a new OmpR-binding site that is lo...
journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1994-03-01 00:00:00