“Soft”-cuticle protein secondary structure as revealed by FT-Raman, ATR FT-IR and CD spectroscopy

Abstract

The nature of the interaction of insect cuticular proteins and chitin is unknown even though about half of the cuticular proteins sequenced thus far share a consensus region that has been predicted to be the site of chitin binding. We previously predicted the preponderance of a β-pleated sheet in the consensus region and proposed its responsibility for the formation of helicoidal cuticle (Iconomidou et al., Insect Biochem. Mol. Biol. 29 (1999) 285). In this study, we examined experimentally the secondary structure of intact and guanidine hydrochloride extracted cuticle and the cuticular protein extract. The studied cuticle came from the larval dorsal abdomen of the lepidopteran Hyalophora cecropia, a classical example of “soft” cuticle. Analysis with FT-Raman, ATR FT-IR and CD spectroscopy indicates that antiparallel β-pleated sheet is the predominant molecular conformation of “soft-cuticle” proteins both in situ in the cuticle and following extraction. It seems that this conformation dictates the modes of chitin–protein interaction in cuticle, in agreement with earlier proposals (Atkins, J. Biosci. 8 (1985) 375).