Biochemical and compositional analyses of recombinant lecithin:cholesterol acyltransferase (LCAT) obtained from a hepatic source

A. F. Ayyobi, Andras G. Lacko, K. Murray, Maya P. Nair, M. Li, H. O.F. Molhuizen, P. H. Pritchard

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Lecithin:cholesterol acyltransferase (LCAT) is an important plasma glycoprotein which plays a central role in lipid metabolism. This protein is responsible for generation of cholesteryl esters in plasma and it has been proposed to play a pivotal role in the reverse cholesterol transport pathway. Structural and functional studies of LCAT have employed various expression systems for production of recombinant LCAT (rLCAT). However, recent studies have shown some differences in the oligosaccharide structure and composition of rLCAT. In this study, we have generated a new hepatic based expression system using McArdle-RH7777 (Mc-7777) cells to produce a recombinant protein most similar to human plasma LCAT. The expressed glycoprotein was compared to the LCAT expressed in previously characterized baby hamster kidney (BHK) cells. Both proteins were compared on the basis of their carbohydrate structure and composition as well as their functional properties. Although the functional properties of both glycoproteins were similar, the carbohydrate structure was significantly different. While BHK-LCAT contained bi-, tri-, and tetraantennary structures, Mc-7777 LCAT presented only biantennary oligosaccharide structures. The difference in glycosylation pattern of rLCAT from Mc-7777 and BHK cells underlines the importance of appropriate expression system, both in vivo and in vitro. (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)1-13
Number of pages13
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume1484
Issue number1
DOIs
StatePublished - 24 Feb 2000

Keywords

  • Carbohydrate structure
  • Enzyme kinetics
  • Lecithin:cholesterol acyltransferase
  • Nuclear magnetic resonance
  • Protein expression
  • Reverse cholesterol transport

Fingerprint Dive into the research topics of 'Biochemical and compositional analyses of recombinant lecithin:cholesterol acyltransferase (LCAT) obtained from a hepatic source'. Together they form a unique fingerprint.

  • Cite this