RT Journal Article
T1 Self-aggregation of a recombinant form of the propeptide NH2-terminal of the precursor of pulmonary surfactant protein SP-B: a conformational study
A1 Bañares-Hidalgo, Ángeles
A1 Bolaños-Gutiérrez, Amada
A1 Pérez Gil, Jesús
A1 Gil Dones, Félix
A1 Estrada Díaz, María Del Pilar
AB A recombinant form of the peptide N-terminally positioned from proSP-B (SP-BN) has been produced in Escherichia coli as fusion with the Maltose Binding Protein, separated from it by Factor Xa cleavage and purified thereafter. This protein module is thought to control assembly of mature SP-B, a protein essential for respiration, in pulmonary surfactant as it progress through the progressively acidified secretory pathway of pneumocytes. Self-aggregation studies of the recombinant propeptide have been carried out as the pH of the medium evolved from neutral to moderately acid, again to neutral and finally basic. The profile of aggregation versus subsequent changes in pH showed differences depending on the ionic strength of the medium, low or moderate, and the presence of additives such as L-arginine (a known aggregation suppressor) and Ficoll 70 (a macromolecular crowder). Circular dichroism studies of SP-BN samples along the aggregation process showed a decrease in α-helical content and a concomitant increase in β-sheet. Intrinsic fluorescence emission of SP-BN was dominated by the emission of Trp residues in neutral medium, being its emission maximum shifted to red at low pH, suggesting that the protein undergoes a pH-dependent conformational change that increases the exposure of their Trp to the environment. A marked increase in the fluorescence emission of the extrinsic probe bis-ANS indicated the exposure of hydrophobic regions of SP-BN at pH 5. The fluorescence of bis-ANS decreased slightly at low ionic strength, but to a great extent at moderate ionic strength when the pH was reversed to neutrality, suggesting that self-aggregation properties of the SP-BN module could be tightly modulated by the conditions of pH and the ionic environment encountered by pulmonary surfactant during assembly and secretion.
PB Oxford University Press
SN 1367-5435
YR 2008
FD 2008
LK https://hdl.handle.net/20.500.14352/94447
UL https://hdl.handle.net/20.500.14352/94447
LA eng
NO Bañares-Hidalgo, A., et al. «Self-Aggregation of a Recombinant Form of the Propeptide NH2-Terminal of the Precursor of Pulmonary Surfactant Protein SP-B: A Conformational Study». Journal of Industrial Microbiology & Biotechnology, vol. 35, n.o 11, noviembre de 2008, pp. 1367-76. https://doi.org/10.1007/s10295-008-0437-3.
NO Research in the laboratory of the authors is funded by grants from Spanish Ministry of Science (BIO2006-03130, CSD2007-00010) and Community of Madrid (P-MAT-000283-0505), and Marie Curie Networks EST-007931 and RTN-512229 from European Commission.
NO Comunidad de Madrid
NO European Commission
NO Ministerio de Ciencia, Innovación y Universidades (España)
DS Docta Complutense
RD 6 abr 2025