Biochemical characteristics of Escherichia coli ATP synthase with insulin peptide A fused to the globular part of the gamma-subunit.
The codon 5383-5385 (CCG) in the atpC gene of the unc operon of Escherichia coli cells was replaced with the sequence encoding peptide A of human insulin. The foreign protein fused to the middle part of the gamma-subunit of ATP synthase affects neither biosynthesis of the chimeric polypeptide nor the integration of the EF(0) x F(1) enzyme into the membranes of the E. coli cells. The inserted peptide A does not inhibit the process of oxidative phosphorylation. The ATPase activity of the mutant EF(0) x F(1) enzyme was equal to that of the wild-type enzyme and was regulated by modifiers in the similar way, suggesting that the space in the stalk area of F(0)/F(1) interaction is enough for the introduction of an additional oligopeptide without changing catalytic properties of the ATP synthase.
Ponomarenko
S V
SV
Institute of Plant Biochemistry of Heinrich Heine University, Düsseldorf, Düsseldorf 40225, Germany. ponomarenko@t-online.de
D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D025261:Mitochondrial Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D007328:Insulin; D011993:Recombinant Fusion Proteins; C474451:F1F0-ATP synthase; D025261:Mitochondrial Proton-Translocating ATPases