Primary structure of cold-adapted alkaline phosphatase from a Vibrio sp. as deduced from the nucleotide gene sequence.
Alkaline phosphatases (AP) are widely distributed in nature, and generally have a dimeric structure. However, there are indications that either monomeric or multimeric bacterial forms may exist. This paper describes the gene sequence of a psychrophilic marine Vibrio AP, previously shown to be particularly heat labile. The kinetic properties were also indicative of cold adaptation. The amino acid sequence of the Vibrio G15-21 AP reveals that the residues involved in the catalytic mechanism, including those ligating the metal ions, have precedence in other characterized APs. Compared with Escherichia coli AP, the two zinc binding sites are identical, whereas the metal binding site, normally occupied by magnesium, is not. Asp-153 and Lys-328 of E. coli AP are His-153 and Trp-328 in Vibrio AP. Two additional stretches of amino acids not present in E. coli AP are found inserted close to the active site of the Vibrio AP. The smaller insert could be accommodated within a dimeric structure, assuming a tertiary structure similar to E. coli AP. In contrast the longer insert would most likely protrude into the interface area, thus preventing dimer formation. This is the first primary structure of a putative monomeric AP, with indications as to the basis for a monomeric existence. Proximity of the large insert loop to the active site may indicate a surrogate role for the second monomer, and may also shape the catalytic as well as stability characteristics of this enzyme.
Asgeirsson
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B
Department of Chemistry, Science Institute, Univerisity of Iceland, Reykjavik. bjarni@raunvis.hi.is
Andrésson
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OS
D000220:Adaptation, Biological; D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D014733:Vibrio / Q000201:enzymology* / Q000235:genetics
D000469:Alkaline Phosphatase