A simple improved method for protein extraction from human head hairs.

This study was conducted to establish a simple efficient method for extracting the protein from human head hair materials, which can be a useful tool for the protein analysis applicable to various types of human head hairs. The method developed saves extraction time and effort considerably. The method includes four steps: cutting the hair samples into small pieces 1-2 mm in length, washing them with distilled water, incubating the hair samples in a buffer solution at 50 degrees C for 24 hr, and finally filtering the incubated mixtures through three layers of nylon mesh. This method is reproducible and reliable. SDS-PAGE analysis of the hair protein extracted by this method shows a clear protein profile on the gel, which is frequently observed in other hair sources. Two smaller sizes of molecular weights are also detected with the SDS-PAGE analysis. Not commonly found in other hair sources, they seem to be other types of human hair proteins.

Molecular and biochemical characterizations of dehydroascorbate reductase from sesame (Sesamum indicum L.) hairy root cultures.

Dehydroascorbate reductase (DHAR) is a biotechnologically or physiologically important reducing enzyme in the ascorbate-glutathione recycling reaction for most higher plants. A DHAR cDNA was isolated from sesame (Sesamum indicum L.) hairy roots, and its structure and biochemical properties were characterized to provide some information about its expressional and biochemical profiles in the hairy root cultures. The cDNA contained a catalytic motif CXXS, which may be indicative of a thiol-dependent redox function. A fusion DHAR expressed in an Escherichia coli expression system was purified with four purification steps until a homogeneous single band signal was seen in an acrylamide gel, and its antibody was prepared for Western blot analyses. The biochemical results showed that the purified recombinant DHAR had an optimal pH of around 6.0, which was different from those (pH 7.8-8.2) of other plant species. The temperature optimal for the DHAR activity was in a relatively wide range of 30-60 degrees C. It was proved by a real-time RT-PCR technique that the transcription activity of the DHAR was about 2-5-fold higher during the first 3 week cultures than during the latter 3 week ones. The highest activity of the sesame DHAR was detected in the 4 week cultures of the hairy roots, after which its activity was rapidly decreased to approximately 80%, suggesting that the most active DHAR occurred in this culture period. Western blot analyses confirmed that the presence of DHAR enzyme was identified in both cultures of the fused E. coli and the sesame hairy roots.