The Plant Plasma Membrane: Structure, Function and Molecular by C. Larsson, I. M. Møller, S. Widell (auth.), Dr. Christer

By C. Larsson, I. M. Møller, S. Widell (auth.), Dr. Christer Larsson, Dr. Ian M. Møller (eds.)

The plasma membrane varieties the residing barrier among the mobilephone and its atmosphere. for that reason it has quite a lot of vital capabilities on the topic of the rules of the composition of the mobilephone inside and to com­ munication with the cellphone external. The plasma membrane has for this reason attracted loads of examine curiosity. until eventually the early 1970's it used to be in basic terms pos­ sible to review the plasma membrane in situ, its constitution e. g. by way of electron microscopy and its functionality e. g. via uptake of radioactively categorized com­ kilos into the intact cellphone or tissue. the 1st isolation of plant protoplasts via enzymatic digestion of the phone wall within the early 1970's was once a tremendous breakthrough in that it supplied direct entry to the outer floor of the plasma membrane. extra importantly, T. okay. Hodges and R. J. Leonard in 1972 released the outline of a style through which a fragment enriched in plasma membranes can be remoted from plant tissues utilizing sucrose gradient centrifugation. hence, the 1970's observed a step forward in our realizing of the structurc and serve as of the plasma membrane. In 1981, S. Widell and C. Larsson released the 1st of a chain of papers within which plasma membrane vesicles of excessive yield and purity have been remoted from quite a lot of plant tissues utilizing aqueous polymer two-phase parti­ tioning.

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Additional info for The Plant Plasma Membrane: Structure, Function and Molecular Biology

Sample text

If so, this may become a very useful marker, since the assay (Avruch and Wallach 1971) is simple and very sensitive, and not least because this activity in animal plasma membrane is located on the outer surface of the s. Widell and C. Larsson 28 membrane. It would therefore provide us with an enzyme marker for the apoplastic surface of the plant plasma membrane which at present is missing (Sect. 3). 3 Membrane Thickness Membrane thickness may be used as a morphological marker for plasma membrane vesicles.

Widell and C. Larsson cannot be used in the presence of vanadate, since vanadate interferes with NADH absorbance. Furthermore, K + and Mg2+ are essential for the activity of the added pyruvate kinase, which means that these ions cannot be omitted. When NADH, phosphoenolpyruvate, pyruvate kinase, and lactate dehydrogenase are omitted, the same assay can be used to measure proton pumping only. The absorbance at 495 nm is then monitored continuously (cf. Chap. 7). Note that the ATPase activity (and not only H+ pumping) with inside-out vesicles is inhibited already at very low Triton concentrations in the coupled assay (Palmgren et al.

1absorbance unit (mg protein)-' min-I, and the corresponding ATPase activity ca. Imol NADH oxidized (ATP consumed) (mg protein)-' min-'. Acridine orange serves as i',. pH indicator, which accumulates in the vesicles where its absorbance is quenched when H+ is pumped across the membrane. Valinomycin and the high concentration of KCl collapse the electrical gradient formed by H+ pumping. BSA binds free fatty acids which can act as uncouplers in the reaction (Palmgren et al. 1990b) and DTT and EDTA keep SH groups intact (for composition of preparation media see Sect.

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