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Isolation and characterization of the rat liver actin N-acetylaminopeptidase
Journal article   Open access   Peer reviewed

Isolation and characterization of the rat liver actin N-acetylaminopeptidase

David R Sheff and Peter A Rubenstein
The Journal of biological chemistry, Vol.267(28), pp.20217-20224
1992
DOI: 10.1016/S0021-9258(19)88689-1
PMID: 1400339
url
https://doi.org/10.1016/S0021-9258(19)88689-1View
Published (Version of record) Open Access

Abstract

ctins 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.
 Fundamental and applied biological sciences. Psychology Biological and medical sciences Hydrolases Analytical, structural and metabolic biochemistry Enzymes and enzyme inhibitors

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