Group specific antibodies against the putative AMP-binding domain signature SGTTGXPKG in peptide synthetases and related enzymes.
The superfamily of adenylate forming enzymes including peptide synthetases, acyl-CoA synthetases and insect luciferases is readily identified by the signature sequence SGTTGXPKG. This sequence including an invariant lysyl residue is located in a disordered loop region and was predicted to be of significant antigenicity. Antibodies were generated employing YTSGTTGRPKGC attached to bovine serum albumin and have been successfully used to identify respective enzymes and adenylate forming domains in multienzyme systems. These include the delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetases of Aspergillus nidulans and Acremonium chrysogenum, gramicidin S synthetase 1 and tyrocidine synthetase 1 from Bacillus brevis, acetyl-CoA synthetase from Alcaligenes eutrophus and a putative peptide synthetase from Metarhizium anisopliae. Weaker or no reactions are observed when the amino acid in position X in the protein is non-basic or hydrophobic, which is respectively the case for gramicidin S synthetase 1 and luciferase.
Etchegaray
A
A
Department of Molecular Biology & Biotechnology, Krebs Institute for Biomolecular Research, University of Sheffield, UK. etchegaray@cena.usp.br
Dieckmann
R
R
Engel
P C
PC
Turner
G
G
von Döhren
H
H
D000249:Adenosine Monophosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D000907:Antibodies, Bacterial; D000937:Antigen-Antibody Reactions; D001419:Bacteria / Q000201:enzymology; D015153:Blotting, Western / Q000379:methods; D002417:Cattle; D003066:Coenzyme A Ligases / Q000032:analysis* / Q000737:chemistry; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000737:chemistry; D008156:Luciferases / Q000032:analysis / Q000737:chemistry; D003904:Mitosporic Fungi / Q000201:enzymology; D009097:Multienzyme Complexes / Q000032:analysis; D009842:Oligopeptides / Q000276:immunology; D010453:Peptide Synthases / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D010456:Peptides, Cyclic / Q000096:biosynthesis; D011817:Rabbits; D012710:Serum Albumin, Bovine
D000596:Amino Acids; D000906:Antibodies; D000907:Antibodies, Bacterial; D000939:Epitopes; D009097:Multienzyme Complexes; D009842:Oligopeptides; D010456:Peptides, Cyclic; D012710:Serum Albumin, Bovine; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D008156:Luciferases; D003066:Coenzyme A Ligases; D010453:Peptide Synthases