A specific hydrophobic core in the alpha-lactalbumin molten globule.

Molten globules are partially structured protein folding intermediates that adopt a native-like overall backbone topology in the absence of extensive detectable tertiary interactions. It is important to determine the extent of specific tertiary structure present in molten globules and to understand the role of specific side-chain packing in stabilizing and specifying molten-globule structure. Previous studies indicate that a small degree of specific side-chain packing stabilizes the structures of the cytochrome c, apomyoglobin, and staphylococcal nuclease molten globules. Here we investigate the extent of specific side-chain packing in the molten globule of alpha-lactalbumin (alpha-LA), a highly fluctuating, non-cooperatively formed molten globule. By analyzing a set of point mutations in the helical domain of alpha-LA, we have identified a stabilizing hydrophobic core. Moreover, this core corresponds to a previously identified structural subdomain and likely contains some native-like packing interactions. Our results suggest that native-like packing of core amino acids helps stabilize molten globules and that some specific interactions can exist in even highly dynamic, fluctuating species.