The proteome of maize leaves: use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints.
As a first step in establishing a proteome database for maize, we have embarked on the identification of the leaf proteins resolved on two-dimensional (2-D) gels. We detected nearly 900 spots on the gels with a pH 4-7 gradient and over 200 spots on the gels with a pH 6-11 gradient when the proteins were visualized with colloidal Coomassie blue. Peptide mass fingerprints for 300 protein spots were obtained with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer and 149 protein spots were identified using the protein databases. We also searched the pdbEST databases to identify the leaf proteins and verified 66% of the protein spots that had been identified using the protein databases. Sixty-seven additional protein spots were identified from expressed sequence tags (ESTs). Many abundant leaf proteins are present in multiple spots. Functions of over 50% of the abundant leaf proteins are either unknown or hypothetical. Our results show that EST databases in conjunction with peptide mass fingerprints can be used for identifying proteins from organisms with incomplete genome sequence information.
Porubleva
L
L
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames 50011, USA.
Vander Velden
K
K
Kothari
S
S
Oliver
D J
DJ
Chitnis
P R
PR
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D018745:Genome, Plant; D010449:Peptide Mapping / Q000379:methods; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D003313:Zea mays / Q000235:genetics / Q000378:metabolism*
D010940:Plant Proteins; D020543:Proteome