Hainantoxin-IV (HNTX-IV) was isolated from the Chinese bird spider Ornithoctorcs hainana and identified as a novel antagonist of tetrodotoxin-sensitive (TTX-S) sodium channels. As revealed by the solution structure of HNTX-IV solved by two-dimensional nuclear magnetic resonance (2D-NMR), HNTX-IV adopts an inhibitor cystine knot motif. To check the role of basic residues during HNTX-IV’s interaction with TTX-S sodium channels, R26A and K27A mutants of HNTX-IV were constructed by solid-phase chemical synthesis. The synthesized peptides were purified and refolded under optimized oxidation conditions. Correct synthesis and folding were confirmed by MALDI-TOF mass spectrometry and NMR spectroscopy, respectively. Using the whole-cell patch-clamp technique, Lys27 but not Arg26 was identified as a key residue for HNTX-IV’s bioactivity against TTX-S sodium channels, because R26A-HNTX-IV showed slightly reduced activity and K27A-HNTX-IV showed almost no inhibition.

Effects of Arg26 and Lys27 mutation on the bioactivity of HNTX-IV

Pituitary gland is critical for the control growth and development in animals. D-galactose induced aging model has been widely used to study the aging mechanism and the screening of drugs. Herein,two dimensional gel electrophoresis ( 2-DE) followed by mass spectrometry( MS) was performed to exemine proteome differences in pituitary gland between control and D-galactose treated mice. Thereafter,pituitary glands were isolated from controlled and D-galactose treated mice,and pituitary gland proteins were subjected to 2-DE based comparative proteomic analysis ( n = 8,4 gels in each group) ,which revealed 46 differentially expressed proteins ( > 2 fold) . Among them,32 were identified successfully by mass spectrometry. The results implicated that the altered proteins were mainly involved in glucose metabolism pathway and might contribute to mitochondrial dysfunction. Our data may help to better understanding of molecular mechanisms of pituitary dysfunction in aging mice induced by Dgalactose.

Differential Proteomic Analysis of Pituitary Gland from D-galactose Induced Aging Mice

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Effects of Arg26 and Lys27 mutation on the bioactivity of HNTX-IV

Differential Proteomic Analysis of Pituitary Gland from D-galactose Induced Aging Mice