Human Adipocyte Lipid-Binding Protein: Purification of the Protein and Cloning of Its Complementary DNA

Cheryl A. Baxa, Ronald S. Sha, Melissa K. Buelt, Anne J. Smith, Valerie Matarese, Laurie L. Chinander, Kyria L. Boundy, David A. Bernlohr

Research output: Contribution to journalArticlepeer-review

88 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)8683-8690
Number of pages8
JournalBiochemistry
Volume28
Issue number22
DOIs
StatePublished - 1989

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