文献类型：期刊文献

英文题名：Elucidation of Xylem-Specific Transcription Factors and Absolute Quantification of Enzymes Regulating Cellulose Biosynthesis in Populus trichocarpa

作者：Loziuk, Philip L.[1] Parker, Jennifer[1] Li, Wei[2] Lin, Chien-Yuan[2] Wang, Jack P.[2] Li, Quanzi[3] Sederoff, Ronald R.[2] Chiang, Vincent L.[2] Muddiman, David C.[1]

第一作者：Loziuk, Philip L.

通信作者：Muddiman, DC[1]

机构：[1]N Carolina State Univ, Dept Chem, WM Keck FTMS Lab, Raleigh, NC 27695 USA;[2]N Carolina State Univ, Dept Forestry & Environm Resources, Forest Biotechnol Grp, Raleigh, NC 27695 USA;[3]Chinese Acad Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China

年份：2015

卷号：14

期号：10

起止页码：4158-4168

外文期刊名：JOURNAL OF PROTEOME RESEARCH

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000362385100007)】；

基金：This material is based upon work supported by the U.S. Department of Energy Office of Science, Office of Science (Biological and Environmental Research) under Award Number DE-SC000691 as well as the NIH/NCSU Molecular Biotechnology Training Program (Grant 5T32GM00-8776-08).

语种：英文

外文关键词：shotgun discovery proteomics; absolute quantification; PC-IDMS; cellulose biosynthesis; transcription factor; targeted mass spectrometiry; SRM

摘要：Cellulose, the main chemical polymer of wood, is the most abundant polysaccharide in nature.(1) The ability to perturb the abundance and structure of cellulose microfibrils is of critical importance to the pulp and paper industry as well as for the textile, wood products, and liquid biofuels industries. Although much has been learned at the transcript level about the biosynthesis of cellulose, a quantitative understanding at the proteome level has yet to be established. The study described herein sought to identify the proteins directly involved in cellulose biosynthesis during wood formation in Populus trichocarpa along with known xylem-specific transcription factors involved in regulating these key proteins. Development of an effective discovery proteomic strategy through a combination of subcellular fractionation of stem differentiating xylem tissue (SDX) with recently optimized FASP digestion protocols, StageTip fractionation, as well as optimized instrument parameters for global proteomic analysis using the quadrupole-orbitrap mass spectrometer resulted in the deepest proteomic coverage of SDX protein from P. trichocarpa with 9,146 protein groups being identified (1% FDR). Of these, 20 cellulosic/hemicellulosic enzymes and 43 xylem-specific transcription factor groups were identified. Finally, selection of surrogate peptides led to an assay for absolute quantification of 14 cellulosic proteins in SDX of P. trichocarpa.