Abstract:
:The lactose-H+ symport protein (LacS) of Streptococcus thermophilus has a carboxyl-terminal regulatory domain (IIALacS) that is homologous to a family of proteins and protein domains of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) in various organisms, of which IIAGlc of Escherichia coli is the best-characterized member. On the basis of these similarities, it was anticipated that IIALacS would be able to perform one or more functions associated with IIAGlc, i.e., carry out phosphoryl transfer and/or affect other catabolic functions. The gene fragment encoding IIALacS was overexpressed in Escherichia coli, and the protein was purified in two steps by metal affinity and anion-exchange chromatography. IIALacS was unable to restore glucose uptake in a IIAGlc-deficient strain, which is consistent with a very low rate of phosphorylation of IIALacS by phosphorylated HPr (HPr approximately P) from E. coli. With HPr approximately P from S. thermophilus, the rate was more than 10-fold higher, but the rate constants for the phosphorylation of IIALacS (k1 = 4.3 x 10(2) M-1 s-1) and dephosphorylation of IIALacS approximately P by HPr (k-1 = 1.1 x 10(3) M-1 s-1) are still at least 4 orders of magnitude lower than for the phosphoryltransfer between IIAGlc and HPr from E. coli. This finding suggests that IIALacS has evolved into a protein domain whose main function is not to transfer phosphoryl groups rapidly. On the basis of sequence alignment of IIA proteins with and without putative phosphoryl transfer functions and the known structure of IIAGlc, we constructed a double mutant [IIALacS(I548E/G556D)] that was predicted to have increased phosphoryl transfer activity. Indeed, the phosphorylation rate of IIALacS(I548E/G556D) by HPr approximately P increased (k1 = 4.0 x 10(3) M-1 s-1) and became nearly independent of the source of HPr approximately P (S. thermophilus, Bacillus subtilis, or E. coli). The increased phosphoryl transfer rate of IIALacS(I548E/G556D) was insufficient to complement IIAGlc in PTS-mediated glucose transport in E. coli. Both IIALacS and IIALacS(I548E/G556D) could replace IIAGlc, but in another function: they inhibited glycerol kinase (inducer exclusion) when present in the unphosphorylated form.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Gunnewijk MG,Postma PW,Poolman Bdoi
10.1128/JB.181.2.632-641.1999subject
Has Abstractpub_date
1999-01-01 00:00:00pages
632-41issue
2eissn
0021-9193issn
1098-5530journal_volume
181pub_type
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01945-14
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abstract::SopE is a bacteriophage-encoded effector protein of Salmonella enterica serovar Typhimurium that is translocated into the cytosol of eukaryotic cells by a type III secretion system (TTSS) (W.-D. Hardt, H. Urlaub, and J. E. Galán, Proc. Natl. Acad. Sci. USA 95:2574-2579, 1998; M. W. Wood, R. Rosqvist, P. B. Mullan, M. ...
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doi:10.1128/jb.185.24.7279-7284.2003
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abstract::Antipain kills Escherichia coli K-12 cells in an exponential manner beginning 1 h after its addition. Mutant strains, delta cya and crp, which are unable to synthesize cyclic adenosine 3',5'-monophosphate (cAMP) and the cAMP receptor protein, respectively, are not affected. Addition of cAMP (5 mM) to antipain-treated ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.139.2.690-693.1979
更新日期:1979-08-01 00:00:00
abstract::The terminal step of triacylglycerol (TAG) formation in the yeast Saccharomyces cerevisiae is catalyzed by the enzyme acyl-CoA:diacylglycerol acyltransferase (DAGAT). In this study we demonstrate that the gene product of YOR245c, Dga1p, catalyzes a major yeast DAGAT activity which is localized to lipid particles. Enzy...
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doi:10.1128/jb.184.2.519-524.2002
更新日期:2002-01-01 00:00:00
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journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.172.12.6981-6990.1990
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abstract::Temperate bacteriophages effect chromosomal evolution of their bacterial hosts, mediating rearrangements and the acquisition of novel genes from other taxa. Although the Haemophilus influenzae genome shows evidence of past phage-mediated lateral transfer, the phages presumed responsible have not been identified. To da...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.24.6893-6905.2002
更新日期:2002-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.144.1.476-479.1980
更新日期:1980-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.162.1.35-41.1985
更新日期:1985-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.162.2.855-857.1985
更新日期:1985-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.93.1.135-141.1967
更新日期:1967-01-01 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:2006-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.181.24.7608-7613.1999
更新日期:1999-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:2003-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.101.1.92-101.1970
更新日期:1970-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.15.4295-4303.2000
更新日期:2000-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.173.10.3231-3234.1991
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
doi:10.1128/jb.171.6.3053-3059.1989
更新日期:1989-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:1996-02-01 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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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.172.11.6291-6299.1990
更新日期:1990-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.188.10.3664-3673.2006
更新日期:2006-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.140.1.37-42.1979
更新日期:1979-10-01 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:1995-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.12.5848-5854.1988
更新日期:1988-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.104.1.581-584.1970
更新日期:1970-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.87.1.33-38.1964
更新日期:1964-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.150.3.1016-1023.1982
更新日期:1982-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.176.1.143-148.1994
更新日期:1994-01-01 00:00:00
abstract::A fatty acid Delta(9)-desaturase mutant of yeast was analyzed to establish the specificity of the fatty acid structural requirement. Several double-bond-containing and substituted fatty acids were tested. It was concluded that the presence of a Delta(9)cis double bond is necessary for growth. The need for a specific c...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.101.1.160-165.1970
更新日期:1970-01-01 00:00:00