title: Isolation and characterization of the rat liver actin N-acetylaminopeptidase.
abstract: Actins from most eukaryotes undergo a unique post-translational modification of the amino terminus called "processing." Processing consists of the removal of an amino-terminal Ac-Met or Ac-Cys to leave an acidic amino-terminal residue. We have previously demonstrated that this reaction is not catalyzed by the ribosomally associated methionine aminopeptidase or by other previously described acetylaminopeptidases. Here we present the isolation and characterization of the actin N-acetylaminopeptidase (ANAP) from rat liver. A five-step purification protocol achieves a 4100-fold purification of the enzyme with an overall 8% recovery of activity. ANAP is a 77-kDa monomer with a pI of 4.6. Using unprocessed yeast actin as a substrate, the Km of ANAP is 3.5 microM. Purified ANAP was used to generate a polyclonal antibody. The antibody has been used along with activity assays to demonstrate the presence of ANAP in a variety of rat tissues. Finally, evidence is presented that in mammals, ANAP may function with a second, as yet unpurified, component to process actin amino termini.
authors.affiliation: Department of Biochemistry, University of Iowa College of Medicine, Iowa City 52242.
authors.forename: D R
authors.initials: DR
authors.lastname: Sheff
authors.forename: P A
authors.initials: PA
authors.lastname: Rubenstein
mesh_terms: D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008099:Liver / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D000199:Actins; D011480:Protease Inhibitors; D000626:Aminopeptidases; C072563:actin N-acetylaminopeptidase