Raman study of the thermal behaviour and conformational stability of basic pancreatic trypsin inhibitor.
We have studied the thermal denaturation of native basic pancreatic trypsin inhibitor (BPTI) by monitoring the Raman bands in the 4000-400 cm(-1) range. In agreement with results obtained by calorimetry, a cooperative melting transition is observed starting at 75 degrees C. This transition is found to involve predominantly the unfolding of helical structures accompanied by beta-aggregation, loss of hydrophobic interactions between side chains and changes in CSSC dihedral angles. However, salt bridge breaking starts near 40 degrees C, as deduced from the nu(s)(COO(-)) band and from the bands close to 1320 and 1345 cm(-1) which for the first time have been shown to be due largely to vibrations of the arginine guanidyl group in BPTI. The thermal stability is, hence, attributable to cooperative contributions from hydrophobic and backbone hydrogen bond interactions as well as from disulfide bonds.
Carmona
Pedro
P
Instituto de Estructura de la Materia (CSIC), Serrano 121, 28006 Madrid, Spain. p.carmona@iem.cfmac.csic.es
Molina
Marina
M
Rodríguez-Casado
Arantxa
A
D000818:Animals; D007611:Aprotinin / Q000737:chemistry*; D002417:Cattle; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D008168:Lung / Q000737:chemistry; D011487:Protein Conformation; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013696:Temperature; D044366:Transition Temperature
D007611:Aprotinin