Expression and complement D activity of porcine adipsin

Citation
Jl. Miner et al., Expression and complement D activity of porcine adipsin, PROT EX PUR, 23(1), 2001, pp. 14-21
Citations number
17
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
PROTEIN EXPRESSION AND PURIFICATION
ISSN journal
1046-5928 → ACNP
Volume
23
Issue
1
Year of publication
2001
Pages
14 - 21
Database
ISI
SICI code
1046-5928(200110)23:1<14:EACDAO>2.0.ZU;2-Z
Abstract
To learn how signals from adipocytes might be involved in regulation of ene rgy intake and storage, we have begun to characterize the porcine complemen t protein, adipsin. Adipsin was originally identified as a protein that is produced rather specifically by adipocytes, is secreted, and is nearly abse nt in several obese rodent models. We now report that porcine adipsin mRNA sequence is 74% identical to rat and predicts a protein that has 82 and 68% identity to human and rat forms, respectively. Porcine adipsin has none of the asparagine glycosylation consensus sites which make recombinant expres sion of mouse adipsin in Escherichia coli impractical. We present a method for engineering the porcine cDNA to facilitate expression by E. coli and pr ovide a protocol for refolding and purifying porcine adipsin protein and fo r immunoassay. We have found that in addition to adipose tissue, adipsin mR NA is present in gut tissues. Coupled with the fact that adipsin is require d for processing of complement C3a-desArg, and that C3a-desArg is a potent stimulant of fatty acid acylation in adipocytes, the production of adipsin in the gut may be related to a mechanism for adipocyte removal of lipid fro m chylomicrons. (C) 2001 Academic Press.