The role of Trp-82 in the folding of intestinal fatty acid binding protein

Paula Dalessio, Susan E. Fromholt, Ira Ropson

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Multiple phases have been observed during the folding and unfolding of intestinal fatty acid binding protein (WT-IFABP) by stopped-flow fluorescence. Site-directed mutagenesis has been used to examine the role of each of the two tryptophans of this protein in these processes. The unfolding and refolding kinetics of the mutant protein containing only tryptophan 82 (W6Y-IFABP) showed that the tryptophan at this location was critical to the fluorescence signal changes observed throughout the unfolding reaction and early in the refolding reaction. However, the kinetic patterns of the mutant protein containing only tryptophan 6 (W82Y-IFABP) indicated that the tryptophan at this location participated in the fluorescence signal changes observed early in the unfolding reaction and late in the refolding reaction. Together, these data suggest that native-like structure was formed first in the vicinity of tryptophan 82, near the center of the hydropliobic core of this β-sheet protein, prior to formation of native-like structure in the periphery of the protein.

Original languageEnglish (US)
Pages (from-to)176-183
Number of pages8
JournalProteins: Structure, Function and Genetics
Volume61
Issue number1
DOIs
StatePublished - Oct 1 2005

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Fatty Acid-Binding Proteins
Tryptophan
Fluorescence
Mutant Proteins
Mutagenesis
Proteins
Kinetics
Site-Directed Mutagenesis

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biochemistry
  • Molecular Biology

Cite this

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abstract = "Multiple phases have been observed during the folding and unfolding of intestinal fatty acid binding protein (WT-IFABP) by stopped-flow fluorescence. Site-directed mutagenesis has been used to examine the role of each of the two tryptophans of this protein in these processes. The unfolding and refolding kinetics of the mutant protein containing only tryptophan 82 (W6Y-IFABP) showed that the tryptophan at this location was critical to the fluorescence signal changes observed throughout the unfolding reaction and early in the refolding reaction. However, the kinetic patterns of the mutant protein containing only tryptophan 6 (W82Y-IFABP) indicated that the tryptophan at this location participated in the fluorescence signal changes observed early in the unfolding reaction and late in the refolding reaction. Together, these data suggest that native-like structure was formed first in the vicinity of tryptophan 82, near the center of the hydropliobic core of this β-sheet protein, prior to formation of native-like structure in the periphery of the protein.",
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The role of Trp-82 in the folding of intestinal fatty acid binding protein. / Dalessio, Paula; Fromholt, Susan E.; Ropson, Ira.

In: Proteins: Structure, Function and Genetics, Vol. 61, No. 1, 01.10.2005, p. 176-183.

Research output: Contribution to journalArticle

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AB - Multiple phases have been observed during the folding and unfolding of intestinal fatty acid binding protein (WT-IFABP) by stopped-flow fluorescence. Site-directed mutagenesis has been used to examine the role of each of the two tryptophans of this protein in these processes. The unfolding and refolding kinetics of the mutant protein containing only tryptophan 82 (W6Y-IFABP) showed that the tryptophan at this location was critical to the fluorescence signal changes observed throughout the unfolding reaction and early in the refolding reaction. However, the kinetic patterns of the mutant protein containing only tryptophan 6 (W82Y-IFABP) indicated that the tryptophan at this location participated in the fluorescence signal changes observed early in the unfolding reaction and late in the refolding reaction. Together, these data suggest that native-like structure was formed first in the vicinity of tryptophan 82, near the center of the hydropliobic core of this β-sheet protein, prior to formation of native-like structure in the periphery of the protein.

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