The action of proteolytic enzymes on chloroplast thylakoid membranes.
Envelope- and stroma-free thylakoid membranes of Vicia faba chloroplasts were incubated with trypsin or pronase for several hours. The indigestible residue was analysed by polyacrylamide gel electrophoresis. Trypsinization resulted in a complete digestion of all proteins with the exception of the pigment-protein complexes as well as a polypeptide not yet characterized. Yet, as compared with untreated material, Complex II was found to have higher electrophoretic mobility. Electron-microscopic studies illustrate that the indigestible residue still has a preserved membrane structure. Disintegration of the thylakoid membranes by sodium dodecyl sulfate followed by trypsinization also resulted in the two complexes while all the other proteins were found to be digested. However, after removal of the lipids the protein moieties of the complexes proved to be easily digestible. From these results it is concluded that pigment-protein interaction may be an important factor in maintaining a conformation rather resistant to perturbants and proteases. In contrast to trypsin, pronase completely digested the polypeptides of the thylakoid membranes including the protein moieties of the pigment-protein complexes leaving an amorphous lipid mass. The results support the assumption that the complexes are necessary to maintain the membrane structure.
D002736:Chloroplasts / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008055:Lipids / Q000032:analysis; D008566:Membranes / Q000378:metabolism* / Q000648:ultrastructure; D010860:Pigments, Biological / Q000032:analysis; D010940:Plant Proteins / Q000032:analysis; D010944:Plants; D011402:Pronase; D011485:Protein Binding; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin
D008055:Lipids; D010860:Pigments, Biological; D010940:Plant Proteins; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin; D011402:Pronase