Comparison of the folding mechanism of highly homologous proteins in the lipid-binding protein family

Ira Ropson, Joshua A. Boyer, Blake A. Schaeffer, Paula Dalessio

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The folding mechanism of two closely related proteins in the intracellular lipid-binding protein family, human bile acid-binding protein (hBABP), and rat bile acid-binding protein (rBABP) were examined. These proteins are 77% identical (93% similar) in sequence. Both of these single domain proteins fit well to a two-state model for unfolding by fluorescence and circular dichroism at equilibrium. Three phases were observed during the unfolding of rBABP by fluorescence but only one phase was observed during the unfolding of hBABP, suggesting that at least two kinetic intermediates accumulate during the unfolding of rBABP that are not observed during the unfolding of hBABP. Fluorine NMR was used to examine the equilibrium unfolding behavior of the W49 side chain in 6-fluorotryptophan-labeled rBABP and hBABP. The structure of rBABP appears to be more dynamic than that of hBABP in the vicinity of W49 in the absence of denaturant, and urea has a greater effect on this dynamic behavior for rBABP than for hBABP. As such, the folding behavior of highly sequence related proteins in this family can be quite different. These differences imply that moderately sized proteins with high sequence and structural similarity can still populate quite different structures during folding.

Original languageEnglish (US)
Pages (from-to)799-806
Number of pages8
JournalProteins: Structure, Function and Bioinformatics
Volume75
Issue number4
DOIs
StatePublished - Jun 1 2009

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Rats
Carrier Proteins
Lipids
Proteins
Fluorescence
Acids
Fluorine
Dichroism
Circular Dichroism
Urea
human AKR1C2 protein
bile acid binding proteins
Nuclear magnetic resonance
Kinetics

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Structural Biology
  • Molecular Biology

Cite this

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title = "Comparison of the folding mechanism of highly homologous proteins in the lipid-binding protein family",
abstract = "The folding mechanism of two closely related proteins in the intracellular lipid-binding protein family, human bile acid-binding protein (hBABP), and rat bile acid-binding protein (rBABP) were examined. These proteins are 77{\%} identical (93{\%} similar) in sequence. Both of these single domain proteins fit well to a two-state model for unfolding by fluorescence and circular dichroism at equilibrium. Three phases were observed during the unfolding of rBABP by fluorescence but only one phase was observed during the unfolding of hBABP, suggesting that at least two kinetic intermediates accumulate during the unfolding of rBABP that are not observed during the unfolding of hBABP. Fluorine NMR was used to examine the equilibrium unfolding behavior of the W49 side chain in 6-fluorotryptophan-labeled rBABP and hBABP. The structure of rBABP appears to be more dynamic than that of hBABP in the vicinity of W49 in the absence of denaturant, and urea has a greater effect on this dynamic behavior for rBABP than for hBABP. As such, the folding behavior of highly sequence related proteins in this family can be quite different. These differences imply that moderately sized proteins with high sequence and structural similarity can still populate quite different structures during folding.",
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Comparison of the folding mechanism of highly homologous proteins in the lipid-binding protein family. / Ropson, Ira; Boyer, Joshua A.; Schaeffer, Blake A.; Dalessio, Paula.

In: Proteins: Structure, Function and Bioinformatics, Vol. 75, No. 4, 01.06.2009, p. 799-806.

Research output: Contribution to journalArticle

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