title: Allosteric Sensing of Fatty Acid Binding by NMR: Application to Human Serum Albumin.
abstract: Human serum albumin (HSA) serves not only as a physiological oncotic pressure regulator and a ligand carrier but also as a biomarker for pathologies ranging from ischemia to diabetes. Moreover, HSA is a biopharmaceutical with a growing repertoire of putative clinical applications from hypovolemia to Alzheimer's disease. A key determinant of the physiological, diagnostic, and therapeutic functions of HSA is the amount of long chain fatty acids (LCFAs) bound to HSA. Here, we propose to utilize (13)C-oleic acid for the NMR-based assessment of albumin-bound LCFA concentration (CONFA). (13)C-Oleic acid primes HSA for a LCFA-dependent allosteric transition that modulates the frequency separation between the two main (13)C NMR peaks of HSA-bound oleic acid (ΔνAB). On the basis of ΔνAB, the overall [(12)C-LCFA]Tot/[HSA]Tot ratio is reproducibly estimated in a manner that is only minimally sensitive to glycation, albumin concentration, or redox potential, unlike other methods to quantify HSA-bound LCFAs such as the albumin-cobalt binding assay.
authors.affiliation: Department of Chemistry and Chemical Biology, and ‡Department of Biochemistry and Biomedical Sciences, McMaster University , 1280 Main Street West, Hamilton, Ontario L8S 4M1, Canada.
authors.forename: Naeimeh
authors.initials: N
authors.lastname: Jafari
authors.affiliation: Department of Chemistry and Chemical Biology, and ‡Department of Biochemistry and Biomedical Sciences, McMaster University , 1280 Main Street West, Hamilton, Ontario L8S 4M1, Canada.
authors.forename: Rashik
authors.initials: R
authors.lastname: Ahmed
authors.affiliation: Department of Chemistry and Chemical Biology, and ‡Department of Biochemistry and Biomedical Sciences, McMaster University , 1280 Main Street West, Hamilton, Ontario L8S 4M1, Canada.
authors.forename: Melanie
authors.initials: M
authors.lastname: Gloyd
authors.affiliation: Department of Chemistry and Chemical Biology, and ‡Department of Biochemistry and Biomedical Sciences, McMaster University , 1280 Main Street West, Hamilton, Ontario L8S 4M1, Canada.
authors.forename: Jonathon
authors.initials: J
authors.lastname: Bloomfield
authors.affiliation: Department of Chemistry and Chemical Biology, and ‡Department of Biochemistry and Biomedical Sciences, McMaster University , 1280 Main Street West, Hamilton, Ontario L8S 4M1, Canada.
authors.forename: Philip
authors.initials: P
authors.lastname: Britz-McKibbin
authors.affiliation: Department of Chemistry and Chemical Biology, and ‡Department of Biochemistry and Biomedical Sciences, McMaster University , 1280 Main Street West, Hamilton, Ontario L8S 4M1, Canada.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Melacini
mesh_terms: D000495:Allosteric Site; D002247:Carbon Isotopes; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009829:Oleic Acids / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D002247:Carbon Isotopes; D009829:Oleic Acids; D012709:Serum Albumin