Human apoC-IV: isolation, characterization, and immunochemical quantification in plasma and plasma lipoproteins.
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.
Kotite
L
L
Cardiovascular Research Institute, University of California-San Francisco, 505 Parnassus Avenue, Rm L-1337, San Francisco, CA 94143, USA.
Zhang
L-H
LH
Yu
Z
Z
Burlingame
A L
AL
Havel
R J
RJ
D020121:5' Untranslated Regions; D000595:Amino Acid Sequence; D000818:Animals; D001056:Apolipoproteins C / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification*; D001483:Base Sequence; D002482:Cellulose / Q000737:chemistry; D002845:Chromatography; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D000882:Haplorhini; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008074:Lipoproteins / Q000737:chemistry; D013058:Mass Spectrometry; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D020033:Protein Isoforms; D011817:Rabbits; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014357:Trypsin / Q000737:chemistry; D014461:Ultracentrifugation
D020121:5' Untranslated Regions; C097349:APOC4 protein, human; C484352:Apoc4 protein, mouse; D001056:Apolipoproteins C; D008074:Lipoproteins; D010455:Peptides; D020033:Protein Isoforms; D002482:Cellulose; D014357:Trypsin