Cloning, structure, and expression of the Escherichia coli K-12 hisC gene

Grisolia, V; Carlomagno, Maria Stella; Nappo, A. G.; Bruni, Carmelo B.

doi:10.1128/jb.164.3.1317-1323.1985

Cited by 34 publications

(9 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…SMUL_1544 displayed an elevated amino acid sequence identity of 46.2%, 46.6%, and 47.4%, respectively, with these proteins. The phylogenetic relationship of this group to the biochemically characterized HisC protein of E. coli K‐12 (GenBank: ; as well as the putative HisC proteins of S. enterica (GenBank: ), or S. multivorans (GenBank: ) is closer compared to the group of CobD‐like decarboxylases. The group of CobD homologs showed a sequence identity of 30–80%, but < 30% to the HisC‐like proteins.…”

Section: Resultsmentioning

confidence: 98%

Structural and functional analysis of an l‐serine O‐phosphate decarboxylase involved in norcobamide biosynthesis

et al. 2019

View full text Add to dashboard Cite

Structural diversity of natural cobamides (Cbas, B12 vitamers) is limited to the nucleotide loop. The loop is connected to the cobalt‐containing corrin ring via an (R)‐1‐aminopropan‐2‐ol O‐2‐phosphate (AP‐P) linker moiety. AP‐P is produced by the l‐threonine O‐3‐phosphate (l‐Thr‐P) decarboxylase CobD. Here, the CobD homolog SMUL_1544 of the organohalide‐respiring epsilonproteobacterium Sulfurospirillum multivorans was characterized as a decarboxylase that produces ethanolamine O‐phosphate (EA‐P) from l‐serine O‐phosphate (l‐Ser‐P). EA‐P is assumed to serve as precursor of the linker moiety of norcobamides that function as cofactors in the respiratory reductive dehalogenase. SMUL_1544 (SmCobD) is a pyridoxal‐5′‐phosphate (PLP)‐containing enzyme. The structural analysis of the SmCobD apoprotein combined with the characterization of truncated mutant proteins uncovered a role of the SmCobD N‐terminus in efficient l‐Ser‐P conversion.

show abstract

Section: Resultsmentioning

confidence: 98%

Structural and functional analysis of an l‐serine O‐phosphate decarboxylase involved in norcobamide biosynthesis

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Culture aliquots were collected and immediately transferred to an equal volume of chilled killer solution (10 mM NaN 3 and 200 lg ml À1 chloramphenicol in diethyl pyrocarbonate-treated water [17]) in order to minimise RNA degradation. The suspension was centrifuged at 5000g for 20 min at 4°C.…”

Section: Rna Extractionmentioning

confidence: 99%

Growth phase-dependent expression of an endoglucanase encoding gene () in A2

Tamburini¹,

Perito²,

Mastromei³

2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

Different growth phases of Streptomyces rochei A2, a cellulolytic strain isolated from termite gut, were determined in liquid culture. The strain exhibited a multiphasic growth curve typical of streptomycetes. In S. rochei A2, the secreted endoglucanase EglS is coded by a TTA-containing gene, conserved in other Streptomyces. The onset of S. rochei A2 eglS gene expression was determined by RT-PCR using total RNA extracted at different growth phases. The eglS gene was expressed only during transition, second rapid growth and stationary phases, the timing of expression being transcriptionally regulated.

show abstract

“…Homologous expression of the E. coli hisC gene in E. coli The hisC gene encoding imidazolacetol-phosphate transaminase (IAPT) was amplified from E. coli K12 chromosomal DNA by PCR. Primers were based on the known sequence of the hisC gene from E. coli (Grisolia et al, 1985). The plasmid pIAPTX99 contained the entire coding region of the hisC gene plus the natural ribosome-binding site cloned downstream of the trc promoter in pTrc99A (Pharmacia).…”

Section: Hind3mentioning

confidence: 99%

Purification and characterization of the imidazoleglycerol-phosphate dehydratase of Saccharomyces cerevisiae from recombinant Escherichia coli

Hawkes

Thomas

Edwards

et al. 1995

Biochemical Journal

View full text Add to dashboard Cite

The HIS3+ gene of Saccharomyces cerevisiae was overexpressed in Escherichia coli and the recombinant imidazoleglycerol-phosphate dehydratase (IGPD) purified to homogeneity. Laser-desorption and electrospray m.s. indicated a molecular ion within 2 units of that expected (23833.3) on the basis of the protein sequence, with about half of the polypeptide lacking the N-terminal formylmethionine residue. IGPD initially purified as an apoprotein was catalytically inactive and mainly a trimer of M(r) 70,000. Addition of Mn2+ (but not Mg2+) caused this to assemble to an active (40 units/mg) enzyme (Mn-IGPD) comprising of 24 subunits (M(r) 573,000) and containing 1.35 +/- 0.1 Mn atoms/polypeptide subunit. An enzyme with an identical activity and metal content was also obtained when the fermenter growth medium of recombinant Escherichia coli was supplemented with MnCl2, and IGPD was purified through as Mn-IGPD rather than as the apoenzyme and assembled in vitro. Inhibition by EDTA indicated that the intrinsic Mn2+ was essential for activity. The retention of activity over time after dilution to very low concentrations of enzyme (< 20 nM) indicated that the metal remained in tight association with the protein. A novel continuous assay method was developed to facilitate the kinetic characterization of Mn-IGPD. At pH 7.0, the Km for IGP was 0.10 +/- 0.02 mM and the Ki value for inhibition by 1,2,4-triazole, 0.12 +/- 0.02 mM. In contrast with other reports, thiols had no influence on catalytic activity. The activity of Mn-IGPD varied with enzyme concentration in such a way as to suggest that it dissociates to a less active form at very low concentrations. Significant inhibition by the product, imidazole acetol phosphate, was inferred from the shape of the progress curve. Titration with, the potent competitive inhibitor, 2-hydroxy-3-(1,2,4-triazol-1-yl)propyl phosphonate indicated that Mn-IGPD contained 0.9 +/- 0.1 catalytic sites/protomer. The activity nearly doubled in the presence of high concentrations of Mn2+; the apparent Ks for stimulation was 20 microM. The basis of this effect was obscure, since there was no corresponding increase in the titre of active sites. Neither was there a discernable shift in the values of Km or Ki (above), although exogenous Mn2+ did reduce the optimum pH for kcat, from 7.2 to 6.8. On the basis of a single site/subunit, the maximum rate of catalytic turnover at 30 degrees C was 32 s-1.

show abstract

Cloning, structure, and expression of the Escherichia coli K-12 hisC gene

Cited by 34 publications

References 58 publications

Structural and functional analysis of an l‐serine O‐phosphate decarboxylase involved in norcobamide biosynthesis

Structural and functional analysis of an l‐serine O‐phosphate decarboxylase involved in norcobamide biosynthesis

Growth phase-dependent expression of an endoglucanase encoding gene () in A2

Purification and characterization of the imidazoleglycerol-phosphate dehydratase of Saccharomyces cerevisiae from recombinant Escherichia coli

Contact Info

Product

Resources

About