Circular Dichroism Studies on C-terminal Zinc Finger Domain of Transcription Factor GATA-2

The C-terminal zinc finger domain of GATA-2 transcription factor from Rattus norvegicus has been expressed and purified to elucidate its secondary structure using circular dichroism spectroscopy. Circular dichroism spectra showed that native GATA-2 C-terminal domain of (Cys)4 type zinc finger has 12% α-helix, 36% β-sheet and 52% random coil content. The estimated structure was compared with predicted structures by sequence based prediction software SOPMA and found to be similar. Furthermore, the effect of pH on the secondary structure of GATA-2 C-finger was examined. This study provides an insight into the understanding of the structure and function of transcription factor GATA-2

Studies on the safety of the consumption of collagen extracted from fish scales in healthy volunteers for a long period: placebo controlled double blind trial

The safety of intake of collagen polypeptides（‘EMFCTR-01’; the commercial name ‘Tenshi no RaRa’） extracted from fish scales for consecutive 12 weeks has been examined by the placebo controlled double blind trial. The participants of 45 healthy volunteers between 30 and 55 years old consist of three groups （15 participants × 3） of a placebo group, a usualamount （collagen ‘EMFCTR-01’ 1.0 g/day） consumption group and a triple-amount （collagen ‘EMFCTR-01’ 3.0 g/day） consumption group. There were no abnormal changes in physical examination, hematological examination, blood biochemical analysis and urinalysis. Changes in electrocardiogram, pulse-wave velocity, muscle stiffness, visceral fat, and fecal conditions were all within the physiological variation. In addition, the physician did not find any adverse events due to collagen ‘EMFCTR-01’ consumption in the reviews of medical history. Hence, the safety of the consumption of collagen‘ EMFCTR-01’ for a long period was confirmed

Studies on the safety of excessive intake of collagen extracted from fish scales in healthy volunteers

The safety of excessive intake of collagen polypeptides （‘EMFCTR-01’; the commercial name ‘Tenshi no RaRa’） extracted from fish scales has been examined by the single group study of 10 healthy volunteers consisting of 4 females and 6 males between 20 and 45 years old. The participants consumed the collagen ‘EMFCTR-01’ of the amount five times as much as usual consumption for 4 consecutive weeks. There were no abnormal changes in physical examination, electrocardiogram, hematological examination, blood biochemical analysis and urinalysis. Although the frequency of the daily defecation was increased during the trial of collagen ‘EMFCTR-01’ consumption, it was considered that the collagen might improve the intestinal environment because there were no changes in the stool output as well as fecal properties. Thus, the safety of collagen‘ EMFCTR-01’ was confirmed

Proceedings of the World Medical Conference

Proceedings of the world medical conferenc

Covalent Structural Changes in Unfolded GroES That Lead to Amyloid Fibril Formation Detected by NMR: INSIGHT INTO INTRINSICALLY DISORDERED PROTEINS*

Co-chaperonin GroES from Escherichia coli works with chaperonin GroEL to mediate the folding reactions of various proteins. However, under specific conditions, i.e. the completely disordered state in guanidine hydrochloride, this molecular chaperone forms amyloid fibrils similar to those observed in various neurodegenerative diseases. Thus, this is a good model system to understand the amyloid fibril formation mechanism of intrinsically disordered proteins. Here, we identified a critical intermediate of GroES in the early stages of this fibril formation using NMR and mass spectroscopy measurements. A covalent rearrangement of the polypeptide bond at Asn45-Gly46 and/or Asn51-Gly52 that eventually yield β-aspartic acids via deamidation of asparagine was observed to precede fibril formation. Mutation of these asparagines to alanines resulted in delayed nucleus formation. Our results indicate that peptide bond rearrangement at Asn-Gly enhances the formation of GroES amyloid fibrils. The finding provides a novel insight into the structural process of amyloid fibril formation from a disordered state, which may be applicable to intrinsically disordered proteins in general