TY - JOUR
T1 - Proteomic analysis of reactive oxygen species (ROS)-related proteins in rice roots
AU - Kim, Sang Gon
AU - Kim, Sun Tae
AU - Kang, Sun Young
AU - Wang, Yiming
AU - Kim, Wook
AU - Kang, Kyu Young
N1 - Funding Information:
Acknowledgments This work was supported by Grant No. CG1122 from the Crop Functional Genomic Center; by a grant from KOSEF/ MOST to the Environmental Biotechnology National Core Research Center (to S.G. Kim and S.T. Kim); and by scholarships from the Brain Korea 21 Program, Ministry of Education and Human Resources Development, Korea (to S.G. Kim, Y. Wang).
PY - 2008/2
Y1 - 2008/2
N2 - To investigate the rice root proteome, we applied the PEG fractionation technique combined with two-dimensional gel electrophoresis which rendered more well-separated protein spots. Out of the 295 chosen proteins, 93 were identified by MALDI-TOF mass spectrometry. The proteins were classified as relating to metabolism (38.7%), reactive oxygen species (ROS)-related proteins (22.5%), protein processing/degradation (8.6%), stress/defense (7.5%), energy (6.5%) and signal transduction (5.4%). The high percentage of ROS-related proteins found in rice root brings us to assess the roles of ROS on rice root growth. Treatment with ROS quenching chemicals such as reduced glutathione (GSH), diphenyleneiodonium (DPI) and ascorbate inhibited root growth dose-dependently. Forty-nine proteins identified were either up- or down-regulated by GSH treatment, of which 14 were ROS-related proteins, such noticeably modulated ones as glutathione-S-transferase (GST), superoxide dismutases (SOD) and l-ascorbate peroxidases. The protein levels of four GSTs (NS4, 8, 56 and 57), three APXs (NS46, 49 and 50) and MnSOD (NS45) were strongly reduced by GSH treatment but slightly reduced by ascorbate and DPI. Ascorbate and DPI strongly inhibited expression levels of a catalase A (NP23) and an APX (NS65) but did not affect APXs (NS46, 49 and 50) protein levels. Northern analysis demonstrated that changes in transcript levels of five genes - GST (NS4), GST (NS43), Mn-SOD (NS45), APX (NS50) and APX (NS46/49) in response to ROS quenching chemicals were coherent with patterns shown in two-dimensional electrophoresis analyses. Taken together, we suggest that these proteins may take part in an important role in maintaining cellular redox homeostasis during rice root growth.
AB - To investigate the rice root proteome, we applied the PEG fractionation technique combined with two-dimensional gel electrophoresis which rendered more well-separated protein spots. Out of the 295 chosen proteins, 93 were identified by MALDI-TOF mass spectrometry. The proteins were classified as relating to metabolism (38.7%), reactive oxygen species (ROS)-related proteins (22.5%), protein processing/degradation (8.6%), stress/defense (7.5%), energy (6.5%) and signal transduction (5.4%). The high percentage of ROS-related proteins found in rice root brings us to assess the roles of ROS on rice root growth. Treatment with ROS quenching chemicals such as reduced glutathione (GSH), diphenyleneiodonium (DPI) and ascorbate inhibited root growth dose-dependently. Forty-nine proteins identified were either up- or down-regulated by GSH treatment, of which 14 were ROS-related proteins, such noticeably modulated ones as glutathione-S-transferase (GST), superoxide dismutases (SOD) and l-ascorbate peroxidases. The protein levels of four GSTs (NS4, 8, 56 and 57), three APXs (NS46, 49 and 50) and MnSOD (NS45) were strongly reduced by GSH treatment but slightly reduced by ascorbate and DPI. Ascorbate and DPI strongly inhibited expression levels of a catalase A (NP23) and an APX (NS65) but did not affect APXs (NS46, 49 and 50) protein levels. Northern analysis demonstrated that changes in transcript levels of five genes - GST (NS4), GST (NS43), Mn-SOD (NS45), APX (NS50) and APX (NS46/49) in response to ROS quenching chemicals were coherent with patterns shown in two-dimensional electrophoresis analyses. Taken together, we suggest that these proteins may take part in an important role in maintaining cellular redox homeostasis during rice root growth.
KW - MALDI-TOF
KW - Proteomics
KW - ROS-related protein
KW - Reactive oxygen species (ROS)
KW - Two-dimensional electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=38349138947&partnerID=8YFLogxK
U2 - 10.1007/s00299-007-0441-5
DO - 10.1007/s00299-007-0441-5
M3 - Article
C2 - 17932678
AN - SCOPUS:38349138947
SN - 0721-7714
VL - 27
SP - 363
EP - 375
JO - Plant Cell Reports
JF - Plant Cell Reports
IS - 2
ER -