Differential Extraction and Protein Sequencing Reveals Major Differences in Patterns of Primary Cell Wall Proteins from Plants

• Published in: The Journal of biological chemistry Vol. 272; no. 25; pp. 15841 - 15848
• Main Authors: Robertson, Duncan, Mitchell, Geoffrey P., Gilroy, John S., Gerrish, Chris, Bolwell, G. Paul, Slabas, Antoni R.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.06.1997; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; AMINO ACID SEQUENCES; ARABIDOPSIS; Cell Wall - chemistry; CELL WALLS; CHEMICAL COMPOSITION; DAUCUS CAROTA; Electrophoresis, Polyacrylamide Gel; EXTRACTION; Fabaceae; LYCOPERSICON ESCULENTUM; MOLECULAR SEQUENCE DATA; Nicotiana; NICOTIANA TABACUM; Peptide Mapping - methods; PHASEOLUS VULGARIS; PLANT PROTEINS; Plant Proteins - chemistry; Plants, Medicinal; Plants, Toxic; PROTEINS; Solanum lycopersicum
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.272.25.15841

The proteins of the primary cell walls of suspension cultured cells of five plant species,Arabidopsis, carrot, French bean, tomato, and tobacco, have been compared. The approach that has been adopted is differential extraction followed by SDS-polyacrylamide gel electrophoresis (PAGE), rather than two-dimensional gel analysis, to facilitate protein sequencing. Whole cells were washed sequentially with the following aqueous solutions, CaCl2, CDTA (cyclohexane diaminotetraacetic acid, DTT (dithiothreitol), NaCl, and borate. SDS-PAGE analysis showed consistent differences between species. From the 233 proteins that were selected for sequencing, 63% gave N-terminal data. This analysis shows that (i) patterns of proteins revealed by SDS-PAGE are strikingly different for all five species, (ii) a large number of these proteins cannot be identified by data base searches indicating that a significant proportion of wall proteins have not been previously described, (iii) the major proteins that can be identified belong to very different classes of proteins, (iv) the majority of proteins found in the extracellular growth media are absent from their respective cell wall extracts, and (v) the results of the extraction process are indicative of higher order structure. It appears that aspects of speciation reside in the complement of extracellular wall proteins. The data represent a protein resource for cell wall studies complementary to EST (expressed sequence tag) and DNA sequencing strategies.