Purification and characterization of a novel ribosome-inactivating protein from seeds of Trichosanthes kirilowii Maxim.
A novel ribosome-inactivating protein, designated Trichosanthrip, was purified from mature seeds of Trichosanthes kirilowii Maxim by cation-exchange and gel-filtration chromatography. Trichosanthrip migrated as a single band in SDS-PAGE, with an apparent molecular mass of approximately 13kDa. The molecular mass of Trichosanthrip was 10,964.617Da as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Trichosanthrip showed N-glycosidase activity on 28 S rRNA and strongly inhibited cell-free protein synthesis, with an IC(50) of 1.6ng/ml. Liquid chromatography-tandem mass spectrometry showed that Trichosanthrip was a novel protein with similar sequence to other proteins present in members of the Cucurbitaceae.
Shu
Shao-Hua
SH
Institute of Medicinal Plant Development, College of Plant Science and Technology, Huazhong Agricultural University, 1# Shizishan Street, Hongshan Borough, Wuhan 430070, PR China.
Xie
Guo-Zhen
GZ
Guo
Xiao-Liang
XL
Wang
Mo
M
D002853:Chromatography, Liquid; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012339:RNA, Ribosomal, 28S / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1 / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012639:Seeds / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D029778:Trichosanthes / Q000737:chemistry*
D010446:Peptide Fragments; D010940:Plant Proteins; D012339:RNA, Ribosomal, 28S; D054789:Ribosome Inactivating Proteins, Type 1