Isolation and characterization of the phage T4 PinA protein, an inhibitor of the ATP-dependent lon protease of Escherichia coli.

The Journal of Biological Chemistry
J J HilliardL D Simon

Abstract

The bacteriophage T4 PinA protein, expression of which leads to inhibition of protein degradation in Escherichia coli cells, has been purified from cells carrying multiple copies of the pinA gene. PinA is a heat-stable protein with a subunit Mr of 18,800 and an isoelectric point of 4.6. Under nondenaturing conditions on a gel filtration column, PinA migrated in two peaks corresponding to a dimer and a tetramer. Purified PinA inhibited ATP-dependent protein degradation by Lon protease in vitro; it did not inhibit the activity of other E. coli ATP-dependent proteases, ClpAP or ClpYQ. Furthermore, PinA did not inhibit ATP-independent proteolysis in E. coli cell extracts. PinA binds with high affinity to Lon protease (Kd approximately 10 nM for dimer binding), and a complex with approximately 1 dimer of PinA per tetramer of Lon protease could be isolated by gel filtration. Lon activity was partially restored upon dilution of the PinA-Lon complex to subnanomolar concentrations, indicating that inhibition was reversible and that PinA did not covalently modify Lon protease. PinA was not cleaved by Lon protease, and heating the Lon-PinA complex at 65 degrees C denatured Lon protease and released active PinA. The properties of PinA in v...Continue Reading

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Citations

May 14, 1999·Current Opinion in Microbiology·S Gottesman
Oct 2, 1998·The EMBO Journal·U Jenal, T Fuchs
Oct 23, 2003·Annual Review of Cell and Developmental Biology·Susan Gottesman
Jan 29, 2005·Chembiochem : a European Journal of Chemical Biology·Michael GrollRobert Huber
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Jul 21, 2006·Research in Microbiology·Virginie TsilibarisLaurence Van Melderen
Oct 1, 2021·FEBS Letters·Francesca Coscia, Jan Löwe
May 1, 2010·Biochimie·Neil D Rawlings

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