title: Backbone and side chain dynamics of uncomplexed human adipocyte and muscle fatty acid-binding proteins.
abstract: Adipocyte lipid-binding protein (A-LBP) and muscle fatty acid-binding protein (M-FABP) are members of a family of small ( approximately 15 kDa) cytosolic proteins that are involved in the metabolism of fatty acids and other lipid-soluble molecules. Although highly homologous (65%) and structurally very similar, A-LBP and M-FABP display distinct ligand binding characteristics. Since ligand binding may be influenced by intrinsic protein dynamical properties, we have characterized the backbone and side chain dynamics of uncomplexed (apo) human A-LBP and M-FABP. Backbone dynamics were characterized by measurements of 15N T1 and T2 values and ¿1H¿-15N NOEs. These data were analyzed using model-free spectral density functions and reduced spectral density mapping. The dynamics of methyl-containing side chains were charaterized by measurements of 2H T1 and T1rho relaxation times of 13C1H22H groups. The 2H relaxation data were analyzed using the model-free approach. For A-LBP, 15N relaxation data were obtained for 111 residues and 2H relaxation data were obtained for 42 methyl groups. For M-FABP, 15N relaxation data were obtained for 111 residues and 2H relaxation data were obtained for 53 methyl groups. The intrinsic flexibilities of these two proteins are compared, with particular emphasis placed on binding pocket residues. There are a number of distinct dynamical differences among corresponding residues between the two proteins. In particular, many residues display greater backbone picosecond to nanosecond and/or microsecond to millisecond time scale mobility in A-LBP relative to M-FABP, including F57, K58, and most residues in alpha-helix 2 (residues 28-35). Variations in the dynamics of this region may play a role in ligand selectivity. The side chains lining the fatty acid binding pocket display a wide range of motional restriction in both proteins. Side chains showing distinct dynamical differences between the two proteins include those of residues 20, 29, and 51. This information provides a necessary benchmark for determining dynamical changes induced by ligand binding and may ultimately lead to an enhanced understanding of ligand affinity and selectivity among fatty acid-binding proteins.
authors.affiliation: Department of Macromolecular NMR, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000, USA.
authors.forename: K L
authors.initials: KL
authors.lastname: Constantine
authors.forename: M S
authors.initials: MS
authors.lastname: Friedrichs
authors.forename: M
authors.initials: M
authors.lastname: Wittekind
authors.forename: H
authors.initials: H
authors.lastname: Jamil
authors.forename: C H
authors.initials: CH
authors.lastname: Chu
authors.forename: R A
authors.initials: RA
authors.lastname: Parker
authors.forename: V
authors.initials: V
authors.lastname: Goldfarb
authors.forename: L
authors.initials: L
authors.lastname: Mueller
authors.forename: B T
authors.initials: BT
authors.lastname: Farmer
mesh_terms: D017667:Adipocytes / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D003198:Computer Simulation; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D006801:Humans; D006859:Hydrogen; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000378:metabolism; D018996:Myelin P2 Protein / Q000737:chemistry* / Q000378:metabolism; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011487:Protein Conformation; D013816:Thermodynamics; D025521:Tumor Suppressor Proteins
chemical_list: D002352:Carrier Proteins; C107608:FABP7 protein, human; C082667:Fabp5 protein, mouse; C086936:Fabp7 protein, mouse; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D018996:Myelin P2 Protein; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D009587:Nitrogen Isotopes; D025521:Tumor Suppressor Proteins; D006859:Hydrogen