caa a22 4500 475744225 CHVBK 20180406123508.0 cr unu---uuuuu 170329e20000401xx s 000 0 eng 10.1023/A:1007011904904 doi (NATIONALLICENCE)springer-10.1023/A:1007011904904 Indentification of Venom Proteins of Spider S. huwena on Two-Dimensional Electrophoresis Gel by N-Terminal Microsequencing and Mass Spectrometric Peptide Mapping [Elektronische Daten] [Songping Liang, Xiaolan Li, Menglin Cao, Jinyun Xie, Ping Chen, Renhuai Huang] Venom proteins of the spider Selenocosmia huwena were separated by two-dimensional gel electrophoresis, with the separation in the first dimension on a wide range of immobilized pH (3-10) gradients. Over 300 protein spots were presented on a silver-stained 2D gel. The protein spots with molecular weight > 10 kDa were analyzed, after electrotransferring to polyvinyldene difluoride (PVDF) membrane, by N-terminal microseqencing. Some of the silver-stained protein spots with molecular weight over 10 kDa were analyzed and identified by employing an improved procedure of mass spectrometric peptide mapping, including (1) in-gel reduction, alkylation, and enzymatic digestion; (2) extraction and desalting by using the pipette tip containing a small C18 microcolumn (Ziptip™); and (3) direct MAIDI-TOF mass analysis and protein database searching. Several known toxins such as HWTX-I, HWTX-II, HWTX-IV, and SHL-I were identified and some new components were found among these protein spots. Plenum Publishing Corporation, 2000 MALDI-TOF mass nationallicence proteome nationallicence spider venom nationallicence 2D-PAGE nationallicence Liang Songping College of Life Science, Hunan Normal University, 410081, Changsha, China aut Li Xiaolan College of Life Science, Hunan Normal University, 410081, Changsha, China aut Cao Menglin College of Life Science, Hunan Normal University, 410081, Changsha, China aut Xie Jinyun College of Life Science, Hunan Normal University, 410081, Changsha, China aut Chen Ping College of Life Science, Hunan Normal University, 410081, Changsha, China aut Huang Renhuai College of Life Science, Hunan Normal University, 410081, Changsha, China aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/3(2000-04-01), 225-229 0277-8033 19:3<225 2000 19 10930 https://doi.org/10.1023/A:1007011904904 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007011904904 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liang Songping College of Life Science, Hunan Normal University, 410081, Changsha, China aut NATIONALLICENCE 700 1- Li Xiaolan College of Life Science, Hunan Normal University, 410081, Changsha, China aut NATIONALLICENCE 700 1- Cao Menglin College of Life Science, Hunan Normal University, 410081, Changsha, China aut NATIONALLICENCE 700 1- Xie Jinyun College of Life Science, Hunan Normal University, 410081, Changsha, China aut NATIONALLICENCE 700 1- Chen Ping College of Life Science, Hunan Normal University, 410081, Changsha, China aut NATIONALLICENCE 700 1- Huang Renhuai College of Life Science, Hunan Normal University, 410081, Changsha, China aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/3(2000-04-01), 225-229 0277-8033 19:3<225 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer