Isolation of plant cell wall proteins.
The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks: (1) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure; (2) polysaccharide networks of cellulose, hemicelluloses, and pectins form potential traps for contaminants such as intracellular proteins; (3) the presence of proteins interacting in many different ways with the polysaccharide matrix require different procedures to elute them from the cell wall. Three categories of CWP are distinguished: labile proteins that have little or no interactions with cell wall components, weakly bound proteins extractable with salts, and strongly bound proteins. Two alternative protocols are decribed for cell wall proteomics: (1) nondestructive techniques allowing the extraction of labile or weakly bound CWP without damaging the plasma membrane; (2) destructive techniques to isolate cell walls from which weakly or strongly bound CWP can be extracted. These protocols give very low levels of contamination by intracellular proteins. Their application should lead to a realistic view of the cell wall proteome at least for labile and weakly bound CWP extractable by salts.
Jamet
Elisabeth
E
UMR 5546 CNRS-Université Paul Sabatier-Toulouse III, Castanet-Tolosan, France.
Boudart
Georges
G
Borderies
Giséle
G
Charmont
Stephane
S
Lafitte
Claude
C
Rossignol
Michel
M
Canut
Herve
H
Pont-Lezica
Rafael
R
D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000302:isolation & purification*; D002473:Cell Wall / Q000737:chemistry*
D029681:Arabidopsis Proteins