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Originally published In Press as doi:10.1194/jlr.M300087-JLR200 on April 16, 2003

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Journal of Lipid Research, Vol. 44, 1387-1394, July 2003
Copyright © 2003 by American Society for Biochemistry and Molecular Biology

Human apoC-IV

: isolation, characterization, and immunochemical quantification in plasma and plasma lipoproteins

L. Kotite*, L-H. Zhang1,*, Z. Yu2,{dagger}, A. L. Burlingame{dagger} and R. J. Havel3,*

* Cardiovascular Research Institute, University of California, San Francisco, 505 Parnassus Avenue, Rm L-1337, San Francisco, CA 94143
{dagger} Department of Pharmaceutical Chemistry, 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: 1776–1783). 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 {rho} < 1.21 g/ml is closely related to plasma triglyceride concentration up to 1,770 mg/dl, varying from 0.1–1.9 mg/dl. Neither the human nor rabbit apoC-IV gene contains a typical TATA box in the 5'-flanking region, but the 5'-untranslated region of the rabbit gene contains a unique purine-rich sequence, GGGACAG(G/A), repeated nine times in tandem, with an additional two within the 5'-flanking sequence.

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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