The microfibrillar proteins of human hair: separation by high-performance liquid chromatography and isolation of some proteins enriched in glycine and tyrosine.

The microfibrillar proteins of human hair have been studied by reversed-phase high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A chromatographic procedure which isolates the microfibrillar proteins from other hair-matrix proteins and separates them into collectable fractions has been introduced. These fibrous proteins fall into two major subgroups which are resolved into six components. The same procedure has also resulted in the identification and simultaneous separation of a group of proteins rich in glycine and tyrosine never before detected in human hair. Comparative electrophoretic studies of the crude microfibrillar proteins reveal five bands with apparent molecular weights of 47,000, 50,000, 53,000, 57,000, and 62,000. The relationship between the electrophoretic bands and the chromatographic fractions is now under investigation.

Cystine-rich proteins of human hair: characterization and structure as revealed by reversed-phase and size-exclusion high-performance liquid chromatography.

The human hair cystine-rich proteins have been separated through the combined use of reversed-phase and size-exclusion chromatography into more than fifty components. These have been grouped, based on molecular weight, into six families of closely related members. The families range in molecular weight from less than 6500 for the low-molecular-weight components to more than 67 000 for the high-molecular-weight components, with average intermediate values for the other families of 8000, 11 500, 15 500 and 19 000. The results also suggest an organized structure of the hair matrix proteins. The combined use of reversed-phase and size-exclusion high-performance liquid chromatography in these studies presents an example where the quaternary structure of a multi-component protein can be largely deduced from its chromatographic behaviour.

Reversed-phase high-performance liquid chromatography of some ultra-heterogeneous and covalently-modified proteins from human hair.

A new procedure for the fractionation of the heterogeneous cystine-rich proteins from human hair, utilizing reversed-phase high-performance liquid chromatography, is described. Of these proteins 27 fractions have been collected and analyzed for amino acid composition. There seems to be little correlation between the elution order and the hydrophobicity of the fraction constituents except for the late-eluting fractions. Based on the elution profiles and amino acid contents, these fractions appear to fall into four families. The effects of alkyl chain length, flow-rate and gradient slope, as well as various additives to the organic modifier on the separation have also been investigated. A low flow-rate (0.4 ml/min) and a shallow gradient were essential for the separation of these proteins as was the use of short alkyl chain (C4) or medium alkyl chain (C8) columns. However, with the C4 column reproducibility and recovery were excellent.