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Journal of Lipid Research, Vol. 44, 1387-1394, July 2003 Human apoC-IV: isolation, characterization, and immunochemical quantification in plasma and plasma lipoproteins
* Cardiovascular Research Institute, University of California, San Francisco, 505 Parnassus Avenue, Rm L-1337, San Francisco, CA 94143
3 To whom correspondence should be addressed. e-mail: havelr{at}itsa.ucsf.edu
Apolipoprotein C-IV (apoC-IV), the newest member of the low-molecular-weight apoC group, has been characterized in blood plasma of rabbits, in which it is a major proline-rich apoC component (Zhang, L-H., L. Kotite, and R. J. Havel. 1996. Identification, characterization, cloning, and expression of apoC-IV, a novel sialoglycoprotein of rabbit plasma lipoproteins. J. Biol. Chem. 271: 17761783). Although the decoded sequence of mouse and human apoC-IV is known, apoC-IV has not been identified in blood plasma from these or other species. Rabbit apoC-IV exists in several sialoforms, and the asialoform has an acidic isoelectric point. We show that apoC-IV is a basic protein in human, monkey, and mouse plasma, present as a minor apoC component of VLDL. Human apoC-IV, isolated from apo VLDL by DEAE-cellulose chromatography and two-dimensional electrophoresis, was identified by microsequencing four tryptic peptides. The protein exhibits two major isoforms; one is N-glycosylated, and both are variably sialylated. In normolipidemic plasma, greater than 80% of the protein is in VLDL (0.7% of total apo VLDL), with most of the remainder in HDL. The concentration of apoC-IV in the plasma and lipoproteins of This sequence, functioning as a GAGA box that has been implicated in the transcription of a number of genes, may explain the higher level of expression of apoC-IV in rabbits.
Abbreviations: MALDI-MS, matrix-assisted laser desorption ionization mass spectrometry; NEPHGE, nonequilibrium pH gradient electrophoresis; PSD, post-source decay; SAA, serum amyloid A; 2-D, two-dimensional; UTR, untranslated region Supplementary key words chromatography electrophoresis mass spectrometry gene structure 5'-untranslated region apolipoprotein C-IV
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