Identification of low abundance polyA-binding proteins in Arabidopsis chloroplast using polyA-affinity column.
Proteins could be well separated and further identified by the use of 2-DE and related techniques. Yet, there are many proteins could not be detected even by more effective dyes because of their inherent low abundance or their low resolution. As a result, polyA-affinity column was used as a method to enrich polyA-binding proteins and then identified by MALDI-TOF-MS. In this study, 23 Arabidopsis chloroplast protein spots coded by 18 genes were identified, and majority of these proteins were classified into three related categories according to their annotations in the Swiss-Prot database, including NAD-, RNA-, and ATP-binding motifs, respectively. The major goal of the present Arabidopsis chloroplast proteomics project was to identify novel polyA-binding proteins or protein isoforms located in Arabidopsis chloroplasts and the specific research of cellular proteins with extremely low transcription levels could be fulfilled.
Ni
Rui-Juan
RJ
Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, 150040, Harbin, China.
Shen
Zhuo
Z
Yang
Chuan-Ping
CP
Wu
Ya-Dan
YD
Bi
Ying-Dong
YD
Wang
Bai-Chen
BC
D017360:Arabidopsis / Q000737:chemistry / Q000378:metabolism*; D002736:Chloroplasts / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D009994:Osmolar Concentration; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D039102:Poly(A)-Binding Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010940:Plant Proteins; D039102:Poly(A)-Binding Proteins; D011061:Poly A