Abstract

Biochemical Characteristics of Escherichia coli ATP Synthase with Insulin Peptide A Fused to the Globular Part of the gamma-Subunit

S. V. Ponomarenko

Institute of Plant Biochemistry of Heinrich Heine University, Düsseldorf, Universitätsstr. 1, 40225 Düsseldorf, Germany; fax: (49) 211-8113706; E-mail: ponomarenko@t-online.de

Received March 15, 2006; Revision received May 8, 2006
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 EF₀·F₁ 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 EF₀·F₁ 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 F₀/F₁ interaction is enough for the introduction of an additional oligopeptide without changing catalytic properties of the ATP synthase.
KEY WORDS: gamma-subunit of ATP synthase, insulin peptide A, chimeric protein expression
DOI: 10.1134/S0006297906090094