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[Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV].
Sheng Wu Gong Cheng Xue Bao. 2005 Jan; 21(1):92-6.SW

Abstract

Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule.

Authors+Show Affiliations

College of Life Science, Hunan Normal University, Changsha 410081, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

15859335

Citation

Xu, Xia, et al. "[Solid-phase Synthesis and Biological Characterization of S12A-HNTX-IV and R29A-HNTX-IV: Two Mutants of Hainantoxin-IV]." Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, vol. 21, no. 1, 2005, pp. 92-6.
Xu X, Xiong X, Li DL, et al. [Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV]. Sheng Wu Gong Cheng Xue Bao. 2005;21(1):92-6.
Xu, X., Xiong, X., Li, D. L., Xiao, Y. C., Wang, X. C., & Liang, S. P. (2005). [Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV]. Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, 21(1), 92-6.
Xu X, et al. [Solid-phase Synthesis and Biological Characterization of S12A-HNTX-IV and R29A-HNTX-IV: Two Mutants of Hainantoxin-IV]. Sheng Wu Gong Cheng Xue Bao. 2005;21(1):92-6. PubMed PMID: 15859335.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV]. AU - Xu,Xia, AU - Xiong,Xia, AU - Li,Dong-Ling, AU - Xiao,Yu-Cheng, AU - Wang,Xian-Chun, AU - Liang,Song-Ping, PY - 2005/4/30/pubmed PY - 2009/7/23/medline PY - 2005/4/30/entrez SP - 92 EP - 6 JF - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JO - Sheng Wu Gong Cheng Xue Bao VL - 21 IS - 1 N2 - Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule. SN - 1000-3061 UR - https://www.unboundmedicine.com/medline/citation/15859335/[Solid_phase_synthesis_and_biological_characterization_of_S12A_HNTX_IV_and_R29A_HNTX_IV:_two_mutants_of_hainantoxin_IV]_ DB - PRIME DP - Unbound Medicine ER -