Molecular cloning and sequence analysis of cDNA encoding lipoprotein lipase of guinea pig.
We have isolated and sequenced cDNA clones covering the entire coding sequence and short flanking regions of guinea pig lipoprotein lipase. The expression cDNA library used was constructed in lambda gt11 with mRNA derived from adipocytes. The deduced amino acid (aa) sequence starts with a stretch of 17 aa interpreted as a leader peptide. The open reading frame continues with 448 aa residues and ends with a TGA stop codon. Combined with previous data this information allows the assignment of domains in the lipase molecule. A likely candidate for the heparin-binding site is a 9-aa stretch containing five positive charges, analogous to the consensus sequence for receptor-binding sites on apolipoproteins E and B. A previously noted homology to pancreatic lipase is extended. Analysis of polyadenylated RNA from several tissues indicated a high level of expression in adipocytes, heart muscle and mammary gland. No lipoprotein lipase mRNA could be detected in liver. Northern blots revealed three major mRNAs with sizes corresponding to 3.8 kb, 3.3 kb and 2.1 kb, respectively. In adipocytes and heart muscle a fourth mRNA, with an estimated size of 4.5 kb, was also detected. Analysis of genomic DNA by Southern blotting indicated a single gene locus coding for lipoprotein lipase. Hence, modification of the primary transcript seems to be involved in the production of the various mRNAs.
Department of Medical Biochemistry, University of Gothenburg, Sweden.
S
S
Enerbäck
H
H
Semb
G
G
Bengtsson-Olivecrona
P
P
Carlsson
M L
ML
Hermansson
T
T
Olivecrona
G
G
Bjursell
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005260:Female; D006168:Guinea Pigs; D008049:Lipase / Q000235:genetics; D008071:Lipoprotein Lipase / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution
D012333:RNA, Messenger; D004247:DNA; D008049:Lipase; D008071:Lipoprotein Lipase