PMID:1998686

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Citation

No author listed (1991) Deduced amino acid sequence of Escherichia coli adenosine deaminase reveals evolutionarily conserved amino acid residues: implications for catalytic function. Biochemistry 30:2273-80

Abstract

The goal of the research reported here is to identify evolutionarily conserved amino acid residues associated with enzymatic deamination of adenosine. To do this, we isolated molecular clones of the Escherichia coli adenosine deaminase gene by functional complementation of adenosine deaminase deficient bacteria and deduced the amino acid sequence of the enzyme from the nucleotide sequence of the gene. Nucleotide sequence analysis revealed the presence of a 996-nucleotide open reading frame encoding a protein of 332 amino acids having a molecular weight of 36,345. The deduced amino acid sequence of the E. coli enzyme has approximately 33% identity with those of the mammalian adenosine deaminases. With conservative amino acid substitutions the overall sequence homology approaches 50%, suggesting that the structures and functions of the mammalian and bacterial enzymes are similar. Additional amino acid sequence analysis revealed specific residues that are conserved among all three adenosine deaminases and four AMP deaminases for which sequence information is currently available. In view of previously published enzymological data and the conserved amino acid residues identified in this study, we propose a model to account for the enzyme-catalyzed hydrolytic deamination of adenosine. Potential catalytic roles are assigned to the conserved His 214, Cys 262, Asp 295, and Asp 296 residues of mammalian adenosine deaminases and the corresponding conserved amino acid residues in bacterial adenosine deaminase and the eukaryotic AMP deaminases.

Links

PubMed

Keywords

Adenosine Deaminase/genetics; Adenosine Deaminase/metabolism; Amino Acid Sequence; Animals; Base Sequence; Biological Evolution; Cloning, Molecular; Escherichia coli/enzymology; Escherichia coli/genetics; Gene Library; Genes, Bacterial; Genetic Complementation Test; Humans; Kinetics; Mice; Molecular Sequence Data; Plasmids; Sequence Homology, Nucleic Acid

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Annotations

Gene product Qualifier GO ID GO term name Evidence Code with/from Aspect Notes Status

add:Gene_Product(s)

GO:0009168

purine ribonucleoside monophosphate biosynthetic process

IGI: Inferred from Genetic Interaction

EcoliWiki:guaA


P

A double mutant of guaA and add cannot grow on minimal media without supplementation with guanosine (or guanine) and DAP. Expression of a cloned add in trans from a plasmid allows the same strain to grow on minimal media containing DAP (see pp. 2275).

complete

add:Gene_Product(s)

GO:0015950

purine nucleotide interconversion

IGI: Inferred from Genetic Interaction

EcoliWiki:guaA


P

A double mutant of guaA and add cannot grow on minimal media without supplementation with guanosine (or guanine) and DAP. Expression of a cloned add in trans from a plasmid allows the same strain to grow on minimal media containing DAP (see pp. 2275).

complete

add:Gene_Product(s)

GO:0006154

adenosine catabolic process

IGI: Inferred from Genetic Interaction

EcoliWiki:guaA


P

A double mutant of guaA and add cannot grow on minimal media without supplementation with guanosine (or guanine) and DAP. Expression of a cloned add in trans from a plasmid allows the same strain to grow on minimal media containing DAP (see pp. 2275).

complete

add:Gene_Product(s)

GO:0032261

purine nucleotide salvage

IGI: Inferred from Genetic Interaction

EcoliWiki:guaA


P

A double mutant of guaA and add cannot grow on minimal media without supplementation with guanosine (or guanine) and DAP. Expression of a cloned add in trans from a plasmid allows the same strain to grow on minimal media containing DAP (see pp. 2275).

complete

add:Gene_Product(s)

GO:0043103

hypoxanthine salvage

IGI: Inferred from Genetic Interaction

EcoliWiki:guaA


P

A double mutant of guaA and add cannot grow on minimal media without supplementation with guanosine (or guanine) and DAP. Expression of a cloned add in trans from a plasmid allows the same strain to grow on minimal media containing DAP (see pp. 2275).

complete

add:Gene_Product(s)

GO:0046101

hypoxanthine biosynthetic process

IGI: Inferred from Genetic Interaction

EcoliWiki:guaA


P

A double mutant of guaA and add cannot grow on minimal media without supplementation with guanosine (or guanine) and DAP. Expression of a cloned add in trans from a plasmid allows the same strain to grow on minimal media containing DAP (see pp. 2275).

complete

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