TY  - JOUR
TI  - Atomic Force Microscopy (AFM) Analysis of Cell Wall Structural Glycoproteins in vitro
AU  - Chen, Yuning 
AU  - Chen, Liwei 
AU  - Kieliszewsk, Marcia J.
AU  - Cannon, Maura C.
VL  - 5
IS  - 14
PY  - 2015
DA  - 2015/07/20
SP  - e1534
C1  - Bio-protocol 2015;5:e1534
DO  - 10.21769/BioProtoc.1534
UR  - https://doi.org/10.21769/BioProtoc.1534
AB  - Hydroxyproline-rich glycoproteins (HRGPs) are major protein components in dicot primary cell walls and generally account for more than 10% of the wall dry weight. As essential members of the HRGP superfamily, extensins (EXTs) presumably function in the cell wall by assembling into positively charged protein scaffolds (Cannon et al., 2008) that direct the proper deposition of other wall polysaccharides, especially pectins, to ensure correct cell wall assembly (Hall and Cannon, 2002; Lamport et al., 2011a). Extensins are recalcitrant to purification as they are rapidly cross-linked into a covalent network after entering the cell wall but there exists a short time window in which newly synthesized extensin monomers can be extracted (Smith et al., 1984; Smith et al., 1986) by salt elution. A detailed protocol for extraction of extensin and other wall structural proteins has been described earlier (Lamport et al., 2011b). The protocol elaborated here provides an approach to studying the self-assembly of extensins and potentially of other cell wall components in vitro using AFM.
KW  - Plant cell wall
KW  - Structural glycoprotein
KW  - Extensin
KW  - Self-assembly
KW  - Atomic force microscope
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False