Phospholipases as Tools to Study the Localization of Phospholipids in Biological Membranes. A Critical Review

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Abstract
After a general introduction into the field, the principles and prerequisites of phospholipid sidedness experiments are reviewed. Since it is of paramount importance that highly purified phospholipases are used, a brief summary is given of the various procedures that are nowadays available to achieve this goal. A characterization of the different enzymes is also included. This is followed by an ample discussion of the possibilities and limitations in the application of those enzymes in phospholipid localization studies. Special emphasis is put on the experimental pitfalls which are inherent to the use of the phospholipases, since it appears that a great deal of the controversial results can be ascribed to the fact that those problems have not always been recognized. In order to appreciate the special features which are offered by these enzymes, a comparison is made with other techniques commonly used for the localization of phospholipids in biological membranes. The results obtained on artificial membranes, the plasma and subcellular membranes of eukaryotic cells, as well as the membranes of microorganisms, are discussed in detail. Whenever relevant, the possible reasons for the conflicting results which have been obtained are indicated.

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