Molecular cloning and sequence of a cDNA coding for bovine lipoprotein lipase.

Lipoprotein lipase (LPL; triacylglycero-protein acylhydrolase, EC 3.1.1.34) was purified from bovine milk. Synthetic oligonucleotides were prepared, based on the amino acid sequences of three peptides obtained from partial digestion of purified LPL, and were used as probes to isolate cDNA clones for LPL mRNA from a bovine mammary gland. One of the clones, pLPL-49R2, contains an insert cDNA (49R2) of about 3.2 kilobases (kb) that hybridizes to all three probes and encodes a polypeptide that includes the NH2-terminal sequence of bovine LPL reported recently [Ben-Avram, C. M., Ben-Zeev, O., Lee, T. D., Hagga, K., Shively, J. E., Goers, J., Pedersen, M. E., Reeve, J. R. & Schotz, M. C. (1986) Proc. Natl. Acad. Sci. USA 83, 4185-4189]. Complete nucleotide sequence analysis revealed that cDNA insert 49R2 contains the entire coding region for LPL as well as a 3' untranslated region of about 1.6 kb. The predicted amino acid sequence indicates that bovine LPL is a hydrophilic protein consisting of 450 amino acids (Mr 50,548) in its unglycosylated form. Blot hybridization analysis of poly(A)+ mRNA from bovine mammary gland demonstrated that there are at least three sizes of LPL mRNAs--3.2, 2.5, and 1.7 kb--with the 2.5-kb mRNA being the most abundant. Restriction endonuclease mapping of other cDNA clones suggested that the variation in mRNA size results from differential utilization of polyadenylylation signals during mRNA processing.