Isolation and cDNA sequence of human postheparin plasma hepatic triglyceride lipase.
Hepatic triglyceride lipase (H-TGL) was isolated from human postheparin plasma by column chromatography on heparin-Sepharose and phenyl-Sepharose and immunoaffinity chromatography with monoclonal antibodies. The purified enzyme had an apparent molecular weight of 65,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an amino-terminal sequence of Leu-Gly-Gln-Ser-Leu-Lys-Pro-Glu. Partial amino acid sequences of seven cyanogen bromide peptides were obtained. A human hepatoma cDNA library was screened with synthetic oligonucleotides derived from the partial protein sequence. The cloned H-TGL cDNA of 1569 nucleotides predicts a mature protein of 477 amino acids plus a leader sequence of 22 amino acids. Blot hybridization analysis of poly(A)+ mRNA with a putative H-TGL cDNA clone gave a single hybridizing band of 1.7 kilobases. The protein contains four consensus N-glycosylation sequences based on the cDNA sequence. Comparison of the enzyme sequence with that of other lipases reveals highly conserved sequences in regions of putative lipid and heparin binding. The carboxyl terminus of H-TGL contains a highly basic sequence which is not reported to be present in rat H-TGL or other members of the lipase gene family.
Martin
G A
GA
Merrell Dow Research Institute, Cincinnati, Ohio 45215.
Busch
S J
SJ
Meredith
G D
GD
Cardin
A D
AD
Blankenship
D T
DT
Mao
S J
SJ
Rechtin
A E
AE
Woods
C W
CW
Racke
M M
MM
Schafer
M P
MP
D000595:Amino Acid Sequence; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes; D005796:Genes; D006493:Heparin; D006801:Humans; D008049:Lipase / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D010446:Peptide Fragments / Q000032:analysis
D010446:Peptide Fragments; D006493:Heparin; D004247:DNA; D008049:Lipase; D004262:DNA Restriction Enzymes; D003488:Cyanogen Bromide