Purification and characterization of acylation stimulating protein from porcine serum.
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 microM). 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.
Department of Animal Science, Biotechnology Center, University of Nebraska, Lincoln, NE 68583-0908, USA.
Hong
H
Zhang
Sheila K
SK
Jacobi
Candice F
CF
Toombs
Katherine H
KH
Cianflone
Natalya
N
Nersesian
Gautam
G
Sarath
Jess L
JL
Miner
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015153:Blotting, Western; D003001:Cloning, Molecular; D015926:Complement C3a / Q000031:analogs & derivatives*; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007106:Immune Sera; D008297:Male; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D016415:Sequence Alignment; D013552:Swine / Q000097:blood*
D001798:Blood Proteins; D018076:DNA, Complementary; D007106:Immune Sera; D011994:Recombinant Proteins; C044541:complement C3a, des-Arg-(77)-; D015926:Complement C3a