title: The cytoplasmic cyclophilin from Azotobacter vinelandii interacts with phosphate acetyltransferase isoforms enhancing their in vitro activity.
abstract: Cyclophilins belong to the peptidyl-prolyl cis/trans isomerase family of enzymes (EC 5.2.1.8), which accelerate protein folding by catalysing the cis/trans isomerisation of proline imidic peptide bonds. In the present study, by a combination of bioinformatics methods, we identify phosphate acetyltransferase isoforms, AvPTA-1 and AvPTA-2, as potential interacting partners of AvPPIB, the cytoplasmic cyclophilin from Azotobacter vinelandii, and demonstrate their physical interaction by co-expression studies. A decrease in AvPPIB PPIase activity, in the presence of AvPTA-1 or AvPTA-2, further confirms each interaction. Phosphate acetyltransferases (EC 2.3.1.8) catalyse the reversible transfer of the acetyl group from acetyl-P to CoA, forming acetyl-CoA and inorganic phosphate. We examined the effect of AvPPIB on the enzymatic activity of both phosphate acetyltransferase isoforms, and noticed an enhancement of the activity, as well as an alteration of the K                         (                            m                         ) of each isoform, for the reaction substrates, indicating a possible function of AvPPIB in phosphate acetyltransferase activity modulation. Although PPIase activity seems not to be essential for these interactions, since AvPPIB(F99A) active site mutant still interacts with both isoforms, it is responsible for the observed phosphate acetyltransferase activity enhancement as AvPPIB(F99A) enhanced to a significantly lower extent the phosphate acetyltransferase activity of both isoforms compared with AvPPIB.
authors.affiliation: Department of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855, Botanikos, Athens, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Dimou
authors.forename: Anastasia
authors.initials: A
authors.lastname: Venieraki
authors.forename: Chrysoula
authors.initials: C
authors.lastname: Zografou
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Katinakis
mesh_terms: D016948:Azotobacter vinelandii / Q000201:enzymology*; D001426:Bacterial Proteins / Q000378:metabolism; D021983:Cyclophilins / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007527:Isoenzymes / Q000378:metabolism; D050505:Mutant Proteins / Q000378:metabolism; D010709:Phosphate Acetyltransferase / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D007527:Isoenzymes; D050505:Mutant Proteins; D020033:Protein Isoforms; C081786:cyclophilin B; D010709:Phosphate Acetyltransferase; D021983:Cyclophilins