Structural studies on the microfibrillar proteins of wool. Interaction between α-helical segments and reassembly of a four-chain structure

Abstract

The alpha-helix-rich particle of Mr 50 200, derived by limited alpha-chymotryptic digestion of the solubilized microfibrillar proteins from wool alpha-keratin, consists mainly of polypeptide-chain segments of Mr 12 500 (fraction ChC) and 25 000 (fraction ChB). The 12 500-Mr segments are of two types (I and II), which are derived from different polypeptide chains of the microfibrillar complex. Each of these type-I and type-II segments partially self-associates in benign solvents to form either dimers or tetramers. When mixed, the two segments show changes in physical properties (alpha-helix content, difference spectra and molecular weight) indicative of complex-formation. The maximum changes occur when the two segments are mixed in an equimolar ratio. Complexes isolated after rapid dialysis of mixtures from 8 M-urea solution were examined by various methods. A tetrameric structure is the main product formed in all cases, and the maximum amount of tetramer is obtained from equimolar mixtures of the type-I and type-II polypeptides. When urea is removed by dialysis from the unfractionated 12 500-Mr segments (fraction ChC) or from the alpha-helix-rich particle itself, a similar complex of Mr 50 000 is formed. The physical properties of these reconstituted entities (alpha-helix content, molecular weight, thermal stability and exposure of tyrosine residues) are similar to those of the original alpha-helix-rich particle. Cross-linking experiments with dimethyl suberimidate are in agreement with a four-chain complex for the reassembled structures. A pair of double-stranded alpha-helices is proposed for the particle, and is considered to be an integral part of the microfibrillar complex in wool alpha-keratin.