Prekeratin biosynthesis in human scalp epidermis.
Analysis of human scalp epidermal prekeratin polypeptides by two-dimensional gel electrophoresis revealed that each of the bands observed in one-dimensional electrophoresis consisted of three to five polypeptides of the same molecular weight but differing in isoelectric points. It was possible to divide the polypeptides into two families, with isoelectric points in the ranges pH 6.0-8.0 and pH 5.0-5.5 respectively. Incorporation of radiolabelled amino acids into freshly excised pieces of scalp epidermis showed that some of the polypeptides had relatively greater contents of glycine and serine than others. Radiolabelled methionine and leucine were, in contrast, incorporated more or less uniformly into all the polypeptides. After incubation with 32P-labelled orthophosphate, relatively more intense labelling by 32P was observed in the higher molecular weight bands of each family. The most basic of the isoelectric variants in each case did not take up phosphate, implying that at least some of the variation in charge was due to different degrees of phosphorylation. Polyadenylated RNA isolated from scalp epidermis was translated in an RNA-dependent reticulocyte haemolysate system followed by immunoprecipitation and electrophoresis. The polypeptides isolated by using anti-(human scalp prekeratin) immunoglobulin G had similar electrophoretic mobilities in sodium dodecyl sulphate/polyacrylamide gels to authentic prekeratin polypeptides, but had different isoelectric properties. This suggested that the products of keratin gene expression undergo post-translational modification.
D000596:Amino Acids / Q000032:analysis; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D004817:Epidermis / Q000378:metabolism*; D006801:Humans; D007633:Keratins / Q000096:biosynthesis* / Q000235:genetics; D010455:Peptides / Q000032:analysis; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D012313:RNA / Q000378:metabolism; D012535:Scalp / Q000378:metabolism
D000596:Amino Acids; D010455:Peptides; D011498:Protein Precursors; C004818:prekeratin; D012313:RNA; D007633:Keratins