Separation and characterization of Arabidopsis thaliana proteins by two-dimensional gel electrophoresis.
Arabidopsis (Arabidopsis thaliana) proteins were isolated from five tissues (leaf, stem, root, seed and callus), and separated by two-dimensional gel electrophoresis (2-DE). 2-DE was carried out by immobilized pH gradient (IPG) in the first dimension, and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension. With the aid of comigrated five-marker proteins, the patterns of 2-DE gels for each tissue were graphically combined by a computer into a single synthetic image for the integrated Arabidopsis protein spots. The protein spot images, altogether 4763, were characterized by both molecular mass and isoelectric point. Partial amino(N)-terminal sequences of 101 protein spots were analyzed by Edman degradation. Fifty seven proteins were partially sequenced and 46 proteins appeared to have blocked N-termini. Deblocking by hydrazine vapor was carried out on 14 proteins and two of them were found to be pyroglutamyl-blocked N-termini. Forty seven new proteins were found by the present investigation.
Kamo
M
M
Research Institute for Biosciences, Science University of Tokyo, Chiba, Japan.
Kawakami
T
T
Miyatake
N
N
Tsugita
A
A
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry* / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D018517:Plant Roots / Q000737:chemistry; D018547:Plant Stems / Q000737:chemistry; D012639:Seeds / Q000737:chemistry; D017385:Sequence Homology
D018076:DNA, Complementary; D010940:Plant Proteins