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Author |
Mizuguchi, M.; Arai, M.; Ke, Y.; Nitta, K.; Kuwajima, K. |
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Title |
Equilibrium and kinetics of the folding of equine lysozyme studied by circular dichroism spectroscopy |
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Journal Article |
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Year |
1998 |
Publication |
Journal of Molecular Biology |
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283 |
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1 |
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265-277 |
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Keywords |
equine lysozyme; protein folding; molten globule; stopped-flow; folding intermediate |
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Abstract |
The equilibrium unfolding and the kinetics of unfolding and refolding of equine lysozyme, a Ca2+-binding protein, were studied by means of circular dichroism spectra in the far and near-ultraviolet regions. The transition curves of the guanidine hydrochloride-induced unfolding measured at 230 nm and 292.5 nm, and for the apo and holo forms of the protein have shown that the unfolding is well represented by a three-state mechanism in which the molten globule state is populated as a stable intermediate. The molten globule state of this protein is more stable and more native-like than that of α-lactalbumin, a homologous protein of equine lysozyme. The kinetic unfolding and refolding of the protein were induced by concentration jumps of the denaturant and measured by stopped-flow circular dichroism. The observed unfolding and refolding curves both agreed well with a single-exponential function. However, in the kinetic refolding reactions below 3 M guanidine hydrochloride, a burst-phase change in the circular dichroism was present, and the burst-phase intermediate in the kinetic refolding is shown to be identical with the molten globule state observed in the equilibrium unfolding. Under a strongly native condition, virtually all the molecules of equine lysozyme transform the structure from the unfolded state into the molten globule, and the subsequent refolding takes place from the molten globule state. The transition state of folding, which may exist between the molten globule and the native states, was characterized by investigating the guanidine hydrochloride concentration-dependence of the rate constants of refolding and unfolding. More than 80% of the hydrophobic surface of the protein is buried in the transition state, so that it is much closer to the native state than to the molten globule in which only 36% of the surface is buried in the interior of the molecule. It is concluded that all the present results are best explained by a sequential model of protein folding, in which the molten globule state is an obligatory folding intermediate on the pathway of folding. |
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refbase @ user @ |
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3990 |
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Kesel, L.; Neil, D.H. |
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Title |
Restraint and handling of animals |
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1998 |
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Clinical Textbook for Veterinary Technicians. 4th ed. |
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1-26 |
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Cited By (since 1996): 1; Export Date: 21 October 2008 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
4523 |
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Luescher, U.A.; McKeown, D.B.; Dean, H. |
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A cross-sectional study on compulsive behaviour (stable vices) in horses |
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Journal Article |
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1998 |
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Equine veterinary journal. Supplement |
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27 |
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14-18 |
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Cited By (since 1996): 22; Export Date: 21 October 2008 |
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Equine Behaviour @ team @ |
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4527 |
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Zohary, D.; Tchernov, E.; Horwitz, L.K. |
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Title |
The role of unconscious selection in the domestication of sheep and goats |
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1998 |
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J Zool |
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245 |
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Equine Behaviour @ team @ Zohary1998 |
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6240 |
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Author |
Whiten, A. |
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Title |
Imitation of the sequential structure of actions by chimpanzees |
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1998 |
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J Comp Psychol |
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11 |
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Equine Behaviour @ team @ Whiten1998 |
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6291 |
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Author |
Breitenmoser, U. |
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Title |
Large predators in the Alps: the fall and rise of man's competitors |
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1998 |
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Biol Conserv |
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83 |
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Equine Behaviour @ team @ Breitenmoser1998 |
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6450 |
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Gese, E.M.; Ruff, R.L. |
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Title |
Howling by coyotes (Canis latrans): variation among social classes, seasons, and pack sizes |
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1998 |
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Can J Zool |
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76 |
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Equine Behaviour @ team @ Gese1998 |
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6462 |
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Author |
Peters, G.; Tembrock, G. |
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Subharmonics, biphonation, and deterministic chaos in mammal vocalizations |
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1998 |
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Bioacoustics |
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9 |
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Equine Behaviour @ team @ Peters1998 |
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6483 |
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Title |
Animal Acoustic Communication: Sound Analysis and Research Methods |
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Book Whole |
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1998 |
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Springer |
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Berlin |
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Call Number |
Equine Behaviour @ team @ ref56 |
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6497 |
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Author |
de VRIES, H.A.N. |
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Title |
Finding a dominance order most consistent with a linear hierarchy: a new procedure and review |
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1998 |
Publication |
Animal Behaviour. |
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Anim. Behav. |
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55 |
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4 |
Pages |
827-843 |
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A procedure for ordering a set of individuals into a linear or near-linear dominance hierarchy is presented. Two criteria are used in a prioritized way in reorganizing the dominance matrix to find an order that is most consistent with a linear hierarchy: first, minimization of the numbers of inconsistencies and, second, minimization of the total strength of the inconsistencies. The linear ordering procedure, which involves an iterative algorithm based on a generalized swapping rule, is feasible for matrices of up to 80 individuals. The procedure can be applied to any dominance matrix, since it does not make any assumptions about the form of the probabilities of winning and losing. The only assumption is the existence of a linear or near-linear hierarchy which can be verified by means of a linearity test. A review of existing ranking methods is presented and these are compared with the proposed method. |
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refbase @ user @ |
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457 |
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