Rapid and Simultaneous Isolation and Identification of Anti-epilepsy Peptide, Scorpion Analgesic Peptide and Anti-neoplastic Peptide from the Venom of Buthus Marthensii Karsch
By using Shim-Pack WCX-1 cation-exchange high-pressure liquid chromatographic column,anti-epilepsy peptide (AEP), scorpion analgesic peptide (SAP) and anti-neoplastic peptide (ANP),can be rapidly and simultaneously isolated and identified from the venom of Buthus marthensii Karsch, and they can be further purified to a homogeneous component by a subsequent Shim-Pack DIOL-300 size-exclusion high-pressure liquid chromatographic column, respectively.
Objective:To construct a new recombinant antitumor peptide Citrostatin,which consists of 2 anti-tumor domains(by different mechanism),and to study its biological activity,in an effort to search for a peptide with double anti-tumor efficiency.
Used Analgesic-antitumor peptide(AGAP) as the expression model of pSYPU-la,constructed the recombine expression vector pSYPU-la-AGAP,transformed it into the E.
Based on the separation information on the venom of Buthus marthensii Karsch provided by Shim-Pack WCX-1 cation-exchange column, these three kinds of active polypeptides were isolated and identified in large scale in a CM Sepharose CL-6B cation-exchange soft-gel chromatographic column, which has the same carboxymethyl cation-exchange group as Shim-Pack WCX-1 high-pressure column.
We have synthesized and characterized a specifically tritium labeled 3H-Tyr3-TT-232 (30 Ci/mmol) to investigate the effect and the fate of this antitumor peptide on human colon tumor cells.
By using Shim-Pack WCX-1 cation-exchange high-pressure liquid chromatographic column,anti-epilepsy peptide (AEP), scorpion analgesic peptide (SAP) and anti-neoplastic peptide (ANP),can be rapidly and simultaneously isolated and identified from the venom of Buthus marthensii Karsch, and they can be further purified to a homogeneous component by a subsequent Shim-Pack DIOL-300 size-exclusion high-pressure liquid chromatographic column, respectively. Based on the separation information on the venom of Buthus m...
Objective:To construct a new recombinant antitumor peptide Citrostatin,which consists of 2 anti-tumor domains(by different mechanism),and to study its biological activity,in an effort to search for a peptide with double anti-tumor efficiency.Methods: Citrostatin,a 48-amino acid peptide,was artificially synthesized.After purified by HPLC,Citrostain were subjected to the following tests separately: inhibition of endothelial cell proliferation,MTT test of cytotoxicity,inhibition of endothelial cell tube format...
Antitumor peptide AP-9(YKLPGHHHHYRP) was synthesized by Fmoc-solid phase peptide synthesis,and the purity of the product was more than 98%.The coupling reaction between the difficult sequence Fmoc-Arg(Pbf)-OH and H-Pro-Trt resin was investigated.The results showed that the coupling reaction could be completed when it was carried out for 6min at 30℃ by microwave irradiation with THF-NMP(1∶1) as the solvent,and the yield could reach to 80.2%.