The intestinal fatty acid binding protein: the role of turns in fast and slow folding processes. 2002

Krishnananda Chattopadhyay, and Shi Zhong, and Syun-Ru Yeh, and Denis L Rousseau, and Carl Frieden
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, Missouri 63110, USA.

The intestinal fatty acid binding protein is one of a family of proteins that are composed of two beta-sheets surrounding a large interior cavity into which the ligand binds. Glycine residues occur in many of the turns between adjacent antiparallel beta-strands. In previous work, the effect of replacing these glycine residues with valine has been examined with stopped flow instrumentation using intrinsic tryptophan fluorescence spectroscopy [Kim and Frieden (1998) Protein Sci. 7, 1821-1828]. To resolve the burst phase missing in the stopped flow measurements, these valine mutants have been reexamined with sub-millisecond continuous flow instrumentation. Some of the glycine residues have also been replaced with proline, and the folding reactions of these proline mutants have been compared with those of their valine counterparts. In all cases, the stability of the protein is decreased, but some turns appear to be more critical for final structure stabilization than others. Surprisingly, the rate constants observed for all the mutants measured by sub-millisecond continuous flow methods are quite similar (1400-3000 s(-1)), and in all the mutants, there is a shift in the fluorescence emission maximum from that of the unfolded protein to lower wavelengths, suggesting some collapse of the unfolded state within 200 micros. In contrast to the rate constants observed for the initial folding events measured by the sub-millisecond continuous flow method, the rate constants for the slower phase observed in the stopped flow instrument vary widely for the different mutants. The latter step appears to be related to side chain stabilization rather than secondary structure formation. It is also shown that the ligand binds tightly only to the native protein and not to any intermediate forms.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008958 Models, Molecular Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures. Molecular Models,Model, Molecular,Molecular Model
D009363 Neoplasm Proteins Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm. Proteins, Neoplasm
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D002352 Carrier Proteins Proteins that bind or transport specific substances in the blood, within the cell, or across cell membranes. Binding Proteins,Carrier Protein,Transport Protein,Transport Proteins,Binding Protein,Protein, Carrier,Proteins, Carrier
D013050 Spectrometry, Fluorescence Measurement of the intensity and quality of fluorescence. Fluorescence Spectrophotometry,Fluorescence Spectroscopy,Spectrofluorometry,Fluorescence Spectrometry,Spectrophotometry, Fluorescence,Spectroscopy, Fluorescence
D016296 Mutagenesis Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS. Mutageneses
D017510 Protein Folding Processes involved in the formation of TERTIARY PROTEIN STRUCTURE. Protein Folding, Globular,Folding, Globular Protein,Folding, Protein,Foldings, Globular Protein,Foldings, Protein,Globular Protein Folding,Globular Protein Foldings,Protein Foldings,Protein Foldings, Globular
D050556 Fatty Acid-Binding Proteins Intracellular proteins that reversibly bind hydrophobic ligands including: saturated and unsaturated FATTY ACIDS; EICOSANOIDS; and RETINOIDS. They are considered a highly conserved and ubiquitously expressed family of proteins that may play a role in the metabolism of LIPIDS. Fatty Acid-Binding Protein,Adipocyte Lipid Binding Protein,Adipocyte-Specific Fatty Acid-Binding Protein,Brain-Type Fatty Acid-Binding Protein,Cytosolic Lipid-Binding Proteins,Fatty Acid-Binding Protein, Cardiac Myocyte,Fatty Acid-Binding Protein, Myocardial,Fatty Acid-Binding Proteins, Adipocyte-Specific,Fatty Acid-Binding Proteins, Brain-Specific,Fatty Acid-Binding Proteins, Cytosolic-Specific,Fatty Acid-Binding Proteins, Intestinal-Specific,Fatty Acid-Binding Proteins, Liver-Specific,Fatty Acid-Binding Proteins, Myocardial-Specific,Fatty Acid-Binding Proteins, Plasma-Membrane Specific,Intestinal Fatty Acid-Binding Protein,Liver Fatty Acid-Binding Protein,Myocardial Fatty Acid-Binding Protein,Plasma Membrane Fatty Acid-Binding Protein,Acid-Binding Protein, Fatty,Adipocyte Specific Fatty Acid Binding Protein,Brain Type Fatty Acid Binding Protein,Cytosolic Lipid Binding Proteins,Fatty Acid Binding Protein,Fatty Acid Binding Protein, Cardiac Myocyte,Fatty Acid Binding Protein, Myocardial,Fatty Acid Binding Proteins,Fatty Acid Binding Proteins, Adipocyte Specific,Fatty Acid Binding Proteins, Brain Specific,Fatty Acid Binding Proteins, Cytosolic Specific,Fatty Acid Binding Proteins, Intestinal Specific,Fatty Acid Binding Proteins, Liver Specific,Fatty Acid Binding Proteins, Myocardial Specific,Fatty Acid Binding Proteins, Plasma Membrane Specific,Intestinal Fatty Acid Binding Protein,Lipid-Binding Proteins, Cytosolic,Liver Fatty Acid Binding Protein,Myocardial Fatty Acid Binding Protein,Plasma Membrane Fatty Acid Binding Protein,Protein, Fatty Acid-Binding

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