Optical characterization of armadillo Acyl-CoA binding protein

• Published in: International journal of biological macromolecules Vol. 31; no. 1; pp. 19 - 27
• Main Authors: Cavagnari, Brian M, Milikowski, Daniel, Haller, Jorge F, Zanek, Marı́a C, Santomé, José A, Ermácora, Mario R
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 20-12-2002
• Subjects: Acyl-CoA binding protein; Amino Acid Sequence; Animals; Armadillos; Base Sequence; Cattle; Circular Dichroism; Diazepam Binding Inhibitor - chemistry; Escherichia coli - metabolism; Fluorescence; Kinetics; Liver - enzymology; Molecular Sequence Data; Protein Binding; Proteins - chemistry; Rats; Spectrometry, Fluorescence; Temperature; Thermodynamics; Ultraviolet Rays; FPLC, fast protein liquid chromatography; p-CoA, palmitoyl-CoA; armACBP, ratACBP, and bovACBP, ACBP from armadillo Harderian glands, rat and cow, respectively; UV, ultraviolet; ACBP, acyl-CoA binding protein; Fluorescence; SDS, sodium dodecyl sulfate; Circular dichroism; NATA, N-acetyl tryptophanamide; PAGE, polyacrylamide gel electrophoresis; NMR, nuclear magnetic resonance; PCR, polymerase chain reaction; Acyl-CoA binding protein; FABP, fatty-acid binding protein; SEC, size exclusion chromatography; GndCl, guanidinium chloride; λ max, wavelength of a spectral maximum; CD, circular dichroism
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/S0141-8130(02)00045-4

Acyl-CoA binding protein (ACBP) and fatty acid binding protein (FABP) are intracellular transporters of activated and free fatty acids, respectively. Unlike other tissues with active lipid metabolism, armadillo Harderian gland contains much more ACBP than FABP. To characterize armadillo ACBP structure and binding properties, we produced it in
Escherichia coli and carried out detailed fluorescence and circular dichroism spectroscopy studies. The
K
D for palmitoyl-CoA, measured directly by fluorescence and rotatory power, was 34±12 and 75±39 nM, respectively. The structure of armadillo ACBP appears to be very similar to that of bovine and rat liver ACBPs.