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Conformational stability of the NH2-Terminal propeptide of the precursor of pulmonary surfactant protein SP-B

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Bañares Hidalgo, Ángeles and Pérez Gil, Jesús and Estrada, Pilar (2016) Conformational stability of the NH2-Terminal propeptide of the precursor of pulmonary surfactant protein SP-B. PLoS ONE, 111 (7). pp. 1-29. ISSN ESSN: 1932-6203

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Official URL: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0158430



Abstract

Assembly of pulmonary surfactant lipid-protein complexes depends on conformational changes coupled with proteolytic maturation of proSP-B, the precursor of pulmonary surfactant protein B (SP-B), along the surfactant biogenesis pathway in pneumocytes. Conformational destabilization of the N-terminal propeptide of proSP-B (SP-BN) triggers exposure of the mature SP-B domain for insertion into surfactant lipids. We have studied the conformational stability during GdmCl- or urea-promoted unfolding of SP-BN with trp fluorescence and circular dichroism spectroscopies. Binding of the intermediate states to bis-ANS suggests their molten globule-like character. ΔG0 H2O was ~ 12.7 kJ mol-1 either with urea or GdmCl. None of the thermal transitions of SP-BN detected by CD correspond to protein unfolding. Differential scanning calorimetry of SP-BN evidenced two endothermic peaks involved in oligomer dissociation as confirmed with 2 M urea. Ionic strength was relevant since at 150 mM NaCl, the process originating the endotherm at the highest temperature was irreversible (Tm2 = 108.5°C) with an activation energy of 703.8 kJ mol-1. At 500 mM NaCl the process became reversible (Tm2 = 114.4°C) and data were fitted to the Non-two States model with two subpeaks. No free thiols in the propeptide could be titrated by DTNB with or without 5.7 M GdmCl, indicating disulfide bonds establishment.


Item Type:Article
Uncontrolled Keywords:Urea; Oligomers; Fluorescence; Sedimentation; Surfactants; Protein structure; Thermodynamics; Disulfide bonds
Subjects:Medical sciences > Biology > Biochemistry
ID Code:43474
Deposited On:21 Jun 2017 10:06
Last Modified:21 Jun 2017 10:59

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