Histidine-Rich Protein as a Possible Origin of Free Amino Acids of Stratum Corneum

SummaryThe origin of free amino acids and/or their metabolites of the stratum corneum was investigated by pulse-chase experiments using hairless mice.Two, 24, 48, and 72 hrs after the animals were pulse-chased with 3H- histidine or 3H-arginine, radioactivity was determined in the following three fractions of the epidermis: the 0.1 N HCIO4 soluble, ethanol soluble fraction (Fr. I); the 0.1N NClO4 soluble, ethanol insoluble fraction (Fr. II); and the 0.1 N HClO4 insoluble, 8M urea soluble fraction (Fr. III). Radioactivity of the epidermal proteins was also determined in SDS-PAGE gels by the gel slicing method.At first, 3H-histidine and/or 3H-arginine were predominantly incorporated into Fr. III, especially into a certain epidermal protein which showed little mobility on SDS-PAGE. Subsequently, 3H, once incorporated into Fr. III, appeared to shift from Fr. III to Fr. II and, simultaneously, from the top band(s) to a band of 32,000 daltons. As a result of amino acid analysis, a protein of Fr. II was considered a histidine-rich protein corresponding to HRP-II of the new born rats of Ball et al. After 48 hrs, as the radioactivities of Fr. III and Fr. II decreased remarkably, Fr. I gradually increased in radioactivity. The major radioactive substances in Fr. I at 72 hrs were identified as 3H-histidine and 3H-urocanic acid when 3H-histidine was used as a tracer, and as 3H-arginine, 3H-ornithine, and 3H-citrulline in the case of 3H-arginine. The amino acid composition of the histidine-rich protein was very similar to that of the free amino acids of the stratum corneum in the hairless mouse, in which some amino acid metabolites were considered as their precursor amino acids.These results strongly suggest that the free amino acids and/or their metabolites of the stratum corneum might be the final products of a degradation of the histidine-rich protein.