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PMID: 9531482 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

The dhnA gene of Escherichia coli encodes a class I fructose bisphosphate aldolase.

The Biochemical journal ·Vol. 331 ( Pt 2) ·1998-04-15 ·Pages 437-45

Thomson GJ, Howlett GJ, Ashcroft AE, Berry A

Abstract

The gene encoding the Escherichia coli Class I fructose-1, 6-bisphosphate aldolase (FBP aldolase) has been cloned and the protein overproduced in high amounts. This gene sequence has previously been identified as encoding an E. coli dehydrin in the GenBanktrade mark database [gene dhnA; entry code U73760; Close and Choi (1996) Submission to GenBanktrade mark]. However, the purified protein overproduced from the dhnA gene shares all its properties with those known for the E. coli Class I FBP aldolase. The protein is an 8-10-mer with a native molecular mass of approx. 340 kDa, each subunit consisting of 349 amino acids. The Class I enzyme shows low sequence identity with other known FBP aldolases, both Class I and Class II (in the order of 20%), which may be reflected by some novel properties of this FBP aldolase. The active-site peptide has been isolated and the Schiff-base-forming lysine residue (Lys236) has been identified by a combination of site-directed mutagenesis, kinetics and electrospray-ionization MS. A second lysine residue (Lys238) has been implicated in substrate binding. The cloning of this gene and the high levels of overexpression obtained will facilitate future structure-function studies.

MeSH Terms
Amino Acid Sequence Binding Sites Borohydrides/pharmacology Chromatography, High Pressure Liquid Cyanogen Bromide Escherichia coli/enzymology,genetics Fructose-Bisphosphate Aldolase/chemistry,genetics,metabolism Gene Expression Lysine/analysis,chemistry Molecular Sequence Data Molecular Weight Mutagenesis, Site-Directed Phylogeny Polymerase Chain Reaction Schiff Bases Sequence Alignment Sequence Analysis
Chemicals
Borohydrides Schiff Bases sodium borohydride Fructose-Bisphosphate Aldolase Lysine Cyanogen Bromide
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Thomson G J
Department of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.
Howlett G J
Ashcroft A E
Berry A
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3310437
Published
1998-04-15
Pages
437-45
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1219373
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1042/bj3310437 PMC Full Text View journal details
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