RGFGIGS is an amino acid sequence required for acetyl coenzyme A binding and activity of human spermidine/spermine N1acetyltransferase.
Polyamine catabolism is rate limited by spermidine/spermine N1-acetyltransferase (SSAT). Although the amino acid sequence of SSAT is known, the substrate binding and catalytic sites are not. The goal of this study was to define the region responsible for acetyl coenzyme A binding. Human SSAT contains a region of 20 amino acids homologous to several microbial antibiotic N-acetyltransferases. The highest homology is represented in the Campylobacter coli streptothricin acetyltransferase sat4 gene, where 16 identical or highly conserved amino acids exist in a 20-residue stretch. The most conserved residues within this region are RGFGIGS beginning at Arg-101 in the human SSAT. Site-directed mutations to Arg-101, Gly-104, and Gly-106 resulted in proteins with no measurable activity. The G102D mutation produced a partially active protein with a decreased affinity for acetyl coenzyme A and with a Km >10-fold that of the wild-type protein. Analysis using the PredictProtein program suggests a common structure among the microbial and eukaryotic N-acetyltransferases in the region corresponding to the RGFGIGS of human SSAT consisting of an alpha-helix usually preceded by a glycine loop. Our data are consistent with the hypothesis that Arg-101 and the proximal glycine loop are necessary for the activity of human SSAT.
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Johns Hopkins Oncology Center Laboratories, Baltimore, Maryland 21231, USA.
Berkey
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Casero
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D000105:Acetyl Coenzyme A / Q000378:metabolism*; D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002167:Campylobacter / Q000201:enzymology / Q000235:genetics; D016384:Consensus Sequence; D017124:Conserved Sequence; D005798:Genes, Bacterial; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D000099228:Diamine N-Acetyltransferase
D000105:Acetyl Coenzyme A; D000123:Acetyltransferases; D009838:Oligodeoxyribonucleotides; D000099228:Diamine N-Acetyltransferase; C044496:streptothricin acetyltransferase