Proteome analysis of the responses of Panax ginseng C. A. Meyer leaves to high light: use of electrospray ionization quadrupole-time of flight mass spectrometry and expressed sequence tag data.
We performed comparative proteomic analyses in order to understand the physiological responses of ginseng (Panax ginseng C. A. Meyer) to high light (HL). As a first step, we analyzed the proteins expressed in ginseng leaves. Proteins extracted from leaves were separated by two-dimensional polyacrylamide gel electrophoresis. Protein spots were identified by tandem mass spectra analysis using electrospray ionization quadrupole-time of flight mass spectrometry (ESI Q-TOF MS). We used a ginseng expressed sequence tag (EST) database as well as a nonredundant protein database from NCBI to identify proteins. Eighty-one proteins were identified using the nr protein database, 51 of which were also verified from the ginseng EST database. An additional 66 proteins were identified only from the ginseng EST database. Proteins that function in energy metabolism, protein stabilization, and protection against oxidative stress were abundant. To understand the light responses of ginseng leaves, we studied time dependent changes in expressed proteins produced by 0-4 h of HL exposure. Six HL-responsive proteins were identified: three proteins were up-regulated (cytosolic small heat-shock protein, cytosolic ascorbate peroxidase, and putative major latex-like protein) and three proteins were down-regulated (Rieske Fe/S protein, putative 3-beta hydroxysteroid dehydrogenase/isomerase-like protein, and oxygen-evolving enhancer-like protein). Our results show that the ginseng EST database combined with ESI Q-TOF MS analysis can be used to identify ginseng proteins and to elucidate the protective mechanism of ginseng against HL induced damage.
Nam
Myung Hee
MH
Seoul Branch, Korea Basic Science Institute, Seoul, Republic of South Korea.
Heo
Eun Joo
EJ
Kim
Jin Young
JY
Kim
Seung Il
SI
Kwon
Kyung-Hoon
KH
Seo
Jong Bok
JB
Kwon
Ohoak
O
Yoo
Jong Shin
JS
Park
Young Mok
YM
D000595:Amino Acid Sequence; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D020224:Expressed Sequence Tags; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D008027:Light; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000502:physiology; D005894:Panax / Q000378:metabolism*; D018515:Plant Leaves / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D015398:Signal Transduction / Q000502:physiology; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010940:Plant Proteins; D020543:Proteome