%0 Journal Article %A Ugalde, U. O. %A Hernandez, A. %A Galindo, I. %A Pitt, D. %A Barnes, J. C. %A Wakley, G. %T Preparation of right-side-out plasma membrane vesicles from Penicillium cyclopium: a critical assessment of markers %D 1992 %J Microbiology, %V 138 %N 10 %P 2205-2212 %@ 1465-2080 %R https://doi.org/10.1099/00221287-138-10-2205 %I Microbiology Society, %X A plasma membrane fraction was obtained by the combined use of differential centrifugation and aqueous polymer two-phase partitioning techniques. Vanadate-inhibited ATPase and glucan synthase activities were highly enriched in this fraction, although the presence of ATPase activity which was not inhibited by vanadate, nitrate, molybdate, anyimycin A or azide was also detected. Other intracellular membrane marker activities were present at very low or undetectable levels. A further separation step using Percoll density gradient centrifugation resulted in the separation of a fraction which exclusively contained vanadate-inhibited ATPase activity, and was enriched with silicotungstic-acid-staining membrane material. Latency tests performed on the plasma membrane markers showed that the membrane vesicles were in the right-side-out orientation. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-138-10-2205