title: Asp residues of βDELSEED-motif are required for peptide binding in the Escherichia coli ATP synthase.
abstract: This study demonstrates the requirement of Asp-380 and Asp-386 in the βDELSEED-motif of Escherichia coli ATP synthase for peptide binding and inhibition. We studied the inhibition profiles of wild-type and mutant E. coli ATP synthase in presence of c-terminal amide bound melittin and melittin related peptide. Melittin and melittin related peptide inhibited wild-type ATPase almost completely while only partial inhibition was observed in single mutations with replacement of Asp to Ala, Gln, or Arg. Additionally, very little or no inhibition occurred among double mutants βD380A/βD386A, βD380Q/βD386Q, or βD380R/βD386R signifying that removal of one Asp residue allows limited peptide binding. Partial or substantial loss of oxidative phosphorylation among double mutants demonstrates the functional requirement of βD380 and βD386 Asp residues. Moreover, abrogation of wild-type E. coli cell growth and normal growth of mutant cells in presence of peptides provides strong evidence for the requirement of βDELSEED-motif Asp residues for peptide binding. It is concluded that while presence of one Asp residue may allow partial peptide binding, both Asp residues, βD380 and βD386, are essential for proper peptide binding and inhibition of ATP synthase.
authors.affiliation: Department of Biochemistry, Kirksville College of Osteopathic Medicine, A. T. Still University of Health Sciences, Kirksville, MO 63501, United States. Electronic address: zahmad@atsu.edu.
authors.forename: Zulfiqar
authors.initials: Z
authors.lastname: Ahmad
authors.affiliation: Department of Biochemistry, Kirksville College of Osteopathic Medicine, A. T. Still University of Health Sciences, Kirksville, MO 63501, United States.
authors.forename: Junior
authors.initials: J
authors.lastname: Tayou
authors.affiliation: Department of Biochemistry, Kirksville College of Osteopathic Medicine, A. T. Still University of Health Sciences, Kirksville, MO 63501, United States.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Laughlin
mesh_terms: D025181:ATP Synthetase Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D005947:Glucose / Q000494:pharmacology; D008555:Melitten / Q000737:chemistry / Q000494:pharmacology; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding / Q000187:drug effects; D013329:Structure-Activity Relationship; D019802:Succinic Acid / Q000494:pharmacology
chemical_list: D050505:Mutant Proteins; D010455:Peptides; D008555:Melitten; D001224:Aspartic Acid; D019802:Succinic Acid; D025181:ATP Synthetase Complexes; D005947:Glucose
keywords: ATPase; Enzyme inhibition; F(1)F(o) ATP synthase; Peptide; βDELSEED