Linked Life Data

7873555

Source:http://linkedlifedata.com/resource/pubmed/id/7873555

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
1995-4-4
pubmed:abstractText
The rat intestinal fatty acid binding protein is an almost all beta-sheet protein that encloses a large interior cavity into which the fatty acid ligand binds. The protein contains neither cysteine nor proline. In a previous report, six site-directed mutants were obtained, each having a single cysteine residue [Jiang, N., & Frieden, C., (1993) Biochemistry 32, 11015-11021] either in a turn or pointed into the cavity. In this report, each mutant has been unfolded in denaturant and modified with 5-iodoacetamido-fluorescein to introduce a large, bulky, and fluorescent group into the protein at a known position. In all cases, fluorescence changes indicated that the modified protein refolded, and circular dichroism measurements suggested that the refolded protein appeared to be mostly beta-sheet. Denaturation curves suggest that for two mutants intermediate structures exist at denaturant concentrations well below the midpoint of the unfolding curve. For each modified, folded protein, one- and two-dimensional 1H NMR spectra were accumulated and compared to the unmodified and wild-type proteins. While the spectra for the modified proteins showed a number of changes in chemical shifts, they were also consistent with folded proteins on the basis of the degree of chemical shift dispersion. Of the six modified mutant proteins, two appear to have the fluorescein group located in the cavity, but only one of these did not bind fatty acid. The remaining modified proteins are capable of ligand binding.(ABSTRACT TRUNCATED AT 250 WORDS)
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
28
pubmed:volume
34
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2724-30
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:7873555-Animals, pubmed-meshheading:7873555-Binding Sites, pubmed-meshheading:7873555-Carrier Proteins, pubmed-meshheading:7873555-Circular Dichroism, pubmed-meshheading:7873555-Cysteine, pubmed-meshheading:7873555-Fatty Acid-Binding Proteins, pubmed-meshheading:7873555-Fatty Acids, pubmed-meshheading:7873555-Fluoresceins, pubmed-meshheading:7873555-Intestines, pubmed-meshheading:7873555-Ligands, pubmed-meshheading:7873555-Magnetic Resonance Spectroscopy, pubmed-meshheading:7873555-Models, Molecular, pubmed-meshheading:7873555-Mutagenesis, Site-Directed, pubmed-meshheading:7873555-Neoplasm Proteins, pubmed-meshheading:7873555-Nerve Tissue Proteins, pubmed-meshheading:7873555-Protein Folding, pubmed-meshheading:7873555-Protein Structure, Secondary, pubmed-meshheading:7873555-Rats, pubmed-meshheading:7873555-Spectrometry, Fluorescence
pubmed:year
1995
pubmed:articleTitle
Intestinal fatty acid binding protein: folding of fluorescein-modified proteins.
pubmed:affiliation
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't