Purification and characterization of acylation stimulating protein from porcine serum

Authors

Hong Zhang, University of Nebraska-LincolnFollow
Sheila K. Jacobi, University of Nebraska - Lincoln
Candace F. Toombs, University of Nebraska - Lincoln
Katherine H. Cianflone, McGill University
Natalya Nersesian, University of Nebraska-LincolnFollow
Gautam Sarath, University of Nebraska - LincolnFollow
Jess L. Miner, University of Nebraska - LincolnFollow

Date of this Version

2002

Comments

Published in Protein Expression and Purification, vol. 25, no. 2 (July 2002), pp. 348–352. Copyright © 2002 Elsevier Science (USA). Used by permission.

Abstract

A method for purifying acylation stimulating protein (ASP) from porcine serum is described. The mRNA encoding ASP was cloned by reverse transcriptase-polymerase chain reaction which predicted a 76 residue peptide. Based on this sequence, we generated antisera to a C-terminal peptide (ASP_1–20) which aided ASP purification. Identity of the purified protein was verified by N-terminal sequencing. The molecular mass of porcine ASP is 8926. Porcine ASP stimulated esterification of fatty acid into triacylglycerol in cultured human cells with potency similar to that of human ASP (twofold at 5 μM). Based on this evidence that ASP exists in porcine blood, and that it has acylation stimulating activity, we propose that ASP may play a role in regulation of energy storage in adipose tissue in the pig.