A comparative proteome and phosphoproteome analysis of differentially regulated proteins during fertilization in the self-incompatible species Solanum chacoense Bitt.
We have used 2-DE for a time-course study of the changes in protein and phosphoprotein expression that occur immediately after fertilization in Solanum chacoense. The phosphorylation status of the detected proteins was determined with three methods: in vivo labeling, immunodetection, and phosphoprotein-specific staining. Using a pI range of 4-7, 262 phosphorylated proteins could be mapped to the 619 proteins detected by Sypro Ruby staining, representing 42% of the total proteins. Among these phosphoproteins, antibodies detected 184 proteins from which 78 were also detected with either of the other two methods (42%). Pro-Q Diamond phosphoprotein stain detected 111 proteins, of which 76 were also detected with either of the other two methods (68%). The 32P in vivo labeling method detected 90 spots from which 78 were also detected with either of other two methods (87%). On comparing before and after fertilization profiles, 38 proteins and phosphoproteins presented a reproducible change in their accumulation profiles. Among these, 24 spots were selected and analyzed by LC-MS/MS using a hybrid quadrupole-TOF (Q-TOF) instrument. Peptide data were searched against publicly available protein and EST databases, and the putative roles of the identified proteins in early fertilization events are discussed.
Vyetrogon
Kateryna
K
Institut de Recherche en Biologie Végétale (IRBV), Département de sciences biologiques, Université de Montréal, Montréal, QC, Canada.
Tebbji
Faiza
F
Olson
Douglas J H
DJ
Ross
Andrew R S
AR
Matton
Daniel P
DP
D005306:Fertilization / Q000502:physiology*; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism*; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D032322:Solanum / Q000235:genetics / Q000378:metabolism*
D010750:Phosphoproteins; D020543:Proteome