The paper by Movileanu et al. 2001 in this issue of The Journal describes a test of the applicability of the substituted-cysteine-accessibility method (SCAM; Akabas et al. 1992; Akabas et al. 1994) to the problem of locating the narrowest region of a channel lumen. Movileanu et al. 2001 use unusually large sulfhydryl-directed reagents to probe the wide bore channel formed by Staphylococcal α-hemolysin. The structure of the membrane-associated form of this protein has been solved to high resolution (Song et al. 1996). It is a mushroom-shaped heptameric complex that inserts in the membrane of a susceptible cell with the wide cap on the extracellular side, or on the cis side of an artificial planar lipid membrane, and with the stem traversing the bilayer (see Figure 1 in Movileanu et al. 2001). The entrance to...
Scam Feels the Pinch
The effect of steric hindrance along the pathway to a cysteine will depend on whether the concentration of reagent in the vicinity of the target cysteine is at equilibrium with the reagent in the medium or is in a steady state. When transfer rates are greater than the local reaction rate, reagent in a pore closed at one end, or reagent added to both sides of a pore open at both ends, will be at equilibrium with reagent in the medium. In contrast, reagent added to one side only of a pore that is open at both ends (the condition here) will be in a steady state. In the steady state, but not in the equilibrium state, the local concentration of reagent will be sensitive to steric hindrance along the pathway.
Arthur Karlin; Scam Feels the Pinch. J Gen Physiol 1 March 2001; 117 (3): 235–238. doi: https://doi.org/10.1085/jgp.117.3.235
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