title: Short-chain peptides identification of scorpion Buthus martensi Karsch venom by employing high orthogonal 2D-HPLC system and tandem mass spectrometry.
abstract: Scorpion venom contains a considerable variety of neurotoxic peptides that can act on ionic channels. Here, we describe an orthogonal 2D-reversed phase/hydrophilic interaction chromatography system (RPLC/HILIC) and use it to separate short-chain peptides from Asian scorpion Buthus martensi Karsch (BmK) venom in a high throughput format. Due to its high orthogonality and efficiency, 18 homogenous peptides were purified and sequence identified by MS/MS with collision-induced dissociation. Among them, four peptides were discovered, which only have evidence at transcript-level, were first purified from crude venom in this study. Two peptides named BmKK2-b and Martentoxin-b were found the new cleaved chains of known BmKK2 and Martentoxin. In addition, two novel peptides named BmKK12 and BmKK16 in this paper were sequenced by de novo MS/MS, which we predict, are members of potassium channel toxin α-KTx 17 subfamily by homology to other known peptides found in the Swiss-Prot protein database.
authors.affiliation: Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Liaoning, China.
authors.forename: Junyan
authors.initials: J
authors.lastname: Xu
authors.forename: Xiuli
authors.initials: X
authors.lastname: Zhang
authors.forename: Zhimou
authors.initials: Z
authors.lastname: Guo
authors.forename: Jingyu
authors.initials: J
authors.lastname: Yan
authors.forename: Long
authors.initials: L
authors.lastname: Yu
authors.forename: Xiuling
authors.initials: X
authors.lastname: Li
authors.forename: Xingya
authors.initials: X
authors.lastname: Xue
authors.forename: Xinmiao
authors.initials: X
authors.lastname: Liang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018118:Chloride Channels / Q000037:antagonists & inhibitors; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000737:chemistry*; D026902:Potassium Channel Blockers / Q000032:analysis / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012604:Scorpion Venoms / Q000737:chemistry*; D012605:Scorpions; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D018118:Chloride Channels; C445369:KK toxin, Buthus martensi; D010455:Peptides; D026902:Potassium Channel Blockers; D012604:Scorpion Venoms; C471035:martentoxin, Buthus martensi