[Chemical synthesis and characterization of R20A-HWTX-I, a mutant of huwentoxin-I with single residue replacement].
Huwentoxin-I (HWTX-I) is a polypeptide neurotoxin purified from the venom of the spider Selenocosmia huwena. R20A-HWTX-I, a mutant of HWTX-I in which the Arg was replaced by Ala, was synthesized on solid support by using Fmoc chemistry. The synthetic mutant was oxidatively renatured in glutathione-containing buffer, and then isolated by reversed phase and specially designed ion-exchange HPLC. The chemical structure of R20A-HWTX-I was confirmed by amino acid analysis, Edman degradation and MALDI-TOF mass analysis. Physiological experiment showed that the replacement of R20 by a decreased the bioactivity of the HWTX-I by 92%, indicating that R20 is a key residue closely related to the bioactivity of the HWTX-I.
Wang
X C
XC
College of Life Science, Hunan Normal University, Changsha.
Liang
S P
SP
Luo
Z M
ZM
D000818:Animals; D051379:Mice; D009154:Mutation; D009498:Neurotoxins / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D015202:Protein Engineering; D030162:Reptilian Proteins; D013111:Spider Venoms / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship
D009498:Neurotoxins; D030162:Reptilian Proteins; D013111:Spider Venoms; C083624:huwentoxin I