In article <2vgj0d$4q6 at mserv1.dl.ac.uk>
Jannie Hofmeyr <jhsh at maties.sun.ac.za> writes:
>I am looking for a recent review of different techniques
>for permeabilising cells. The most recent one I could find was by Felix
>[Anal. Biochem. 120, 211-234, 1982]. I am specifically interested in
>permeabilising E. coli cells for the purpose of investigating serine
>biosynthesis (no leakage of proteins, permeable to metabolites up to a
>Mr of about 800, NAD(H), ATP, etc.).
Jannie, good to know that you are going to do experients on permeabilised
cells. Not yet physiological but much closer to in vivo. I hope that even with
big holes in the membrane you will find that very little protein leaks out...
I don't know if this helps but Jim Clegg has a few papers on permeabilised
mouse L-929 cells (the methodology might not be applicable to E. coli but you
may pick up useful hints). Look up the following:
Clegg & Jackson (1988) "Glycolysis in permeabilized L-929 cells" Biochem. J.
255, 335-344.
Jackson, Thomson & Clegg (1990) "Glycolysis compared in intact, permeabilized
and sonicated L-929 cells" FEBS Lett. 262, 212-214.
Clegg & Jackson (1990) "Glucose metabolism and the channeling of glycolytic
intermediates in permeabilized L-929 cells" Arch. Biochem. Biophys. 278,
452-460.
Clegg, Jackson &Fendl (1990) "Effects of reduced cell volume and water content
on glycolysis in L-929 cells" J. Cell. Physiol. 142, 386-391.
Ridsdale & Clegg (1991) "Evidence for cooperativity of protein dissolution in
Brij 58 permeabilized L929 cells" J. Cell. Physiol. 147, 242-247.
Hope this helps
Pedro Mendes
prm at aber.ac.uk
