Human adipocyte lipid-binding protein: purification of the protein and cloning of its complementary DNA.
Human adipocyte lipid-binding protein (H-ALBP) was purified from normal subcutaneous adipose tissue to greater than 98% homogeneity, utilizing a combination of acid fractionation, gel filtration, covalent chromatography on activated thiol-Sepharose 4B, and anion-exchange chromatography. Human ALBP comprised about 1% of total cytosolic protein in human adipose tissue, had a relative molecular mass of about 15 kDa, and existed as a monomer in solution. The amino terminus of H-ALBP was blocked to sequencing. When a liposome ligand delivery assay was used, H-ALBP saturably bound oleic acid with about 1 mol of ligand bound per mole of protein. Additionally, H-ALBP saturably bound retinoic acid as determined by the quenching of intrinsic tryptophan fluorescence. A full-length H-ALBP cDNA has been cloned; the sequence predicts a 649-base mRNA comprised of a 62-base 5'-noncoding region containing an 18S ribosome-binding site, a single 396-base open-reading frame, and a 191-base 3'-noncoding region. Comparative sequence analysis indicated that the 132 amino acid H-ALBP is a member of a multigene family of intracellular lipid-binding proteins and contains the consensus substrate phosphorylation sequence for tyrosyl kinases.
Baxa
C A
CA
Department of Biochemistry, University of Minnesota, St. Paul 55108.
Sha
R S
RS
Buelt
M K
MK
Smith
A J
AJ
Matarese
V
V
Chinander
L L
LL
Boundy
K L
KL
Bernlohr
D A
DA
D000273:Adipose Tissue / Q000032:analysis* / Q000166:cytology; D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D019301:Oleic Acid; D009829:Oleic Acids / Q000493:pharmacokinetics; D012313:RNA / Q000302:isolation & purification; D014212:Tretinoin / Q000493:pharmacokinetics; D025521:Tumor Suppressor Proteins
D002352:Carrier Proteins; C107608:FABP7 protein, human; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D009829:Oleic Acids; D025521:Tumor Suppressor Proteins; D019301:Oleic Acid; D014212:Tretinoin; D012313:RNA; D004247:DNA