Probing the antiprotease activity of lambdaCIII, an inhibitor of the Escherichia coli metalloprotease HflB (FtsH).
The CIII protein encoded by the temperate coliphage lambda acts as an inhibitor of the ubiquitous Escherichia coli metalloprotease HflB (FtsH). This inhibition results in the stabilization of transcription factor lambdaCII, thereby helping the phage to lysogenize the host bacterium. LambdaCIII, a small (54-residue) protein of unknown structure, also protects sigma(32), another specific substrate of HflB. In order to understand the details of the inhibitory mechanism of CIII, we cloned and expressed the protein with an N-terminal six-histidine tag. We also synthesized and studied a 28-amino-acid peptide, CIIIC, encompassing the central 14 to 41 residues of CIII that exhibited antiproteolytic activity. Our studies show that CIII exists as a dimer under native conditions, aided by an intersubunit disulfide bond, which is dispensable for dimerization. Unlike CIII, CIIIC resists digestion by HflB. While CIII binds to HflB, it does not bind to CII. On the basis of these results, we discuss various mechanisms for the antiproteolytic activity of CIII.
Halder
Sabyasachi
S
Department of Biochemistry, Bose Institute, P-1/12, CIT Scheme VIIM, Kolkata 700054, India.
Datta
Ajit Bikram
AB
Parrack
Pradeep
P
D049069:ATP-Dependent Proteases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010582:Bacteriophage lambda / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D021122:Protein Subunits; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*
D029968:Escherichia coli Proteins; D021122:Protein Subunits; D014157:Transcription Factors; D014764:Viral Proteins; C054615:cIII protein, Bacteriophage lambda; D049069:ATP-Dependent Proteases; C513383:FtsH protein, E coli