Biochemical characterization of two Azotobacter vinelandii FKBPs and analysis of their interaction with the small subunit of carbamoyl phosphate synthetase.

dc.contributor.author	Dimou, M	en
dc.contributor.author	Zografou, C	en
dc.contributor.author	Venieraki, A	en
dc.contributor.author	Katinakis, P	en
dc.date.accessioned	2014-06-06T06:51:39Z
dc.date.available	2014-06-06T06:51:39Z
dc.date.issued	2012	en
dc.identifier.issn	15734978	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/5622
dc.relation.uri	http://www.scopus.com/inward/record.url?eid=2-s2.0-84875808857&partnerID=40&md5=190dfc758a6848c4203c15bcec1f1ef5	en
dc.subject.other	alanine	en
dc.subject.other	bacterial protein	en
dc.subject.other	carbamoyl phosphate synthase (glutamine hydrolysing)	en
dc.subject.other	chaperone	en
dc.subject.other	citrate synthase	en
dc.subject.other	fk 506 binding protein	en
dc.subject.other	leucine	en
dc.subject.other	phenylalanine	en
dc.subject.other	recombinant protein	en
dc.subject.other	amino acid sequence	en
dc.subject.other	amino acid substitution	en
dc.subject.other	article	en
dc.subject.other	Azotobacter vinelandii	en
dc.subject.other	chemistry	en
dc.subject.other	enzyme active site	en
dc.subject.other	enzyme specificity	en
dc.subject.other	enzymology	en
dc.subject.other	gene expression	en
dc.subject.other	gene expression regulation	en
dc.subject.other	genetics	en
dc.subject.other	growth, development and aging	en
dc.subject.other	hydrolysis	en
dc.subject.other	kinetics	en
dc.subject.other	metabolism	en
dc.subject.other	molecular genetics	en
dc.subject.other	protein analysis	en
dc.subject.other	protein binding	en
dc.subject.other	protein subunit	en
dc.subject.other	site directed mutagenesis	en
dc.subject.other	Alanine	en
dc.subject.other	Amino Acid Sequence	en
dc.subject.other	Amino Acid Substitution	en
dc.subject.other	Azotobacter vinelandii	en
dc.subject.other	Bacterial Proteins	en
dc.subject.other	Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)	en
dc.subject.other	Catalytic Domain	en
dc.subject.other	Citrate (si)-Synthase	en
dc.subject.other	Gene Expression	en
dc.subject.other	Gene Expression Regulation, Bacterial	en
dc.subject.other	Gene Expression Regulation, Enzymologic	en
dc.subject.other	Hydrolysis	en
dc.subject.other	Kinetics	en
dc.subject.other	Leucine	en
dc.subject.other	Molecular Chaperones	en
dc.subject.other	Molecular Sequence Data	en
dc.subject.other	Mutagenesis, Site-Directed	en
dc.subject.other	Phenylalanine	en
dc.subject.other	Protein Binding	en
dc.subject.other	Protein Interaction Mapping	en
dc.subject.other	Protein Subunits	en
dc.subject.other	Recombinant Proteins	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Tacrolimus Binding Proteins	en
dc.title	Biochemical characterization of two Azotobacter vinelandii FKBPs and analysis of their interaction with the small subunit of carbamoyl phosphate synthetase.	en
heal.type	journalArticle	en
heal.publicationDate	2012	en
heal.abstract	FK-506 binding proteins (FKBPs) belong to the peptidyl-prolyl cis/trans isomerase superfamily (PPIases, EC: 5.2.1.8) which catalyzes the interconversion of peptidyl-prolyl bonds while they can also act on polypeptides, as folding helper enzymes. Here, we biochemically characterize two recombinant FKBPs, AvfkbA1 and AvfkbA2, from the soil nitrogen-fixing bacterium Azotobacter vinelandii and show that both possess PPIase activity while AvfkbA2 possesses chaperone activity as well. Further, we demonstrate their physical interaction with AvcarA, the small subunit of carbamoyl phosphate synthetase. Using RT-qPCR, we show that AvfkbA1 and AvfkbA2 are co-expressed with AvcarA under the same growth conditions. A decrease in AvfkbA1 or AvfkbA2 PPIase activity, in the presence of AvcarA, further confirms each interaction. However, PPIase activity does not seem to be essential for these interactions since PPIase active site mutations of both FKBPs do not abolish the AvcarA binding. The P(358) residue of AvcarA, possibly retaining a cis configuration, is critical only for the interaction with AvfkbA1. The presence of either of the two FKBPs did not influence the measured glutamine hydrolyzing activity of AvcarA. Taken together, these data indicate that although the two FKBPs have a common biological substrate they probably have differing physiological roles.	en
heal.journalName	Molecular biology reports	en
dc.identifier.issue	12	en
dc.identifier.volume	39	en
dc.identifier.spage	10003	en
dc.identifier.epage	10012	en