Substituted cysteine accessibility

Karlin, A. and Akabas, M. H. (1998) Substituted-cysteine accessibility method. Methods Enzymol. 293,123-145. 

Several laboratories have replaced consecutive amino acids with cysteine residues as part of a technique called the substituted cysteine accessibility method (SCAM) (Akabas et al., 1992). In this technique, which is described elsewhere in this volume, the accessibihty of the inserted cysteine residues to an aqueous environment is measured by their accessibihty to covalent chemical modifying agents. The effects of the addition of these bulky agents can then be used to explore protein functional domains such as those that line ion channels or ligand binding sites. 

Javitch JA, Shi L, Liapkis G. 2002. Use of the substituted cysteine accessibility method to study the structure and function of G protein-coupled receptors. Methods Enzymol 343 137-156. 

Reeves DC, Goren EN, Akabas MH, Lummis SCR. 2001. Structural and electrostatic properties of the 5-HT3 receptor pore revealed by substituted cysteine accessibility mutagenesis. J Biol Chem 276 42035-42042. 

Klein, H., Garneau, L., Banderali, U., Simoes, M., Parent, L., and Sauve, R. 2007. Structural determinants of the closed KCa3.1 channel pore in relation to channel gating Results from a substituted cysteine accessibility analysis. J. Gen. Physiol. 129 299-315. 

Bogdanov, M., Zhang, W., Xie, J. and Dowhan, W., Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM(TM)) apphcation to lipid-specific membrane protein topogenesis. Methods 36 (2005) 148-171. 

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