title: Thermodynamics, kinetics, and salt dependence of folding of YopM, a large leucine-rich repeat protein.
abstract: Small globular proteins have many contacts between residues that are distant in primary sequence. These contacts create a complex network between sequence-distant segments of secondary structure, which may be expected to promote the cooperative folding of globular proteins. Although repeat proteins, which are composed of tandem modular units, lack sequence-distant contacts, several of considerable length have been shown to undergo cooperative two-state folding. To explore the limits of cooperativity in repeat proteins, we have studied the unfolding of YopM, a leucine-rich repeat (LRR) protein of over 400 residues. Despite its large size and modular architecture (15 repeats), YopM equilibrium unfolding is highly cooperative, and shows a very strong dependence on the concentration of urea. In contrast, kinetic studies of YopM folding indicate a mechanism that includes one or more transient intermediates. The urea dependence of the folding and unfolding rates suggests a relatively small transition state ensemble. As with the urea dependence, we have found an extreme dependence of the free energy of unfolding on the concentration of salt. This salt dependence likely results from general screening of a large number of unfavorable columbic interactions in the folded state, rather than from specific cation binding.
authors.affiliation: T.C. Jenkins Department of Biophysics, Johns Hopkins University, 3400 N. Charles St., Baltimore, MD 21218, USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Kloss
authors.forename: Doug
authors.initials: D
authors.lastname: Barrick
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D007700:Kinetics; D000090263:Leucine-Rich Repeat Proteins; D008958:Models, Molecular; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics; D014364:Tryptophan; D014508:Urea / Q000494:pharmacology
chemical_list: D001425:Bacterial Outer Membrane Proteins; D000090263:Leucine-Rich Repeat Proteins; D011506:Proteins; D012996:Solutions; C060772:yopM protein, Yersinia; D012965:Sodium Chloride; D014364:Tryptophan; D014508:Urea
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