Functional and mutational analysis of the light-harvesting chlorophyll a/b protein of thylakoid membranes.
The precursor for a Lemna light-harvesting chlorophyll a/b protein (pLHCP) has been synthesized in vitro from a single member of the nuclear LHCP multigene family. We report the sequence of this gene. When incubated with Lemna chloroplasts, the pLHCP is imported and processed into several polypeptides, and the mature form is assembled into the light-harvesting complex of photosystem II (LHC II). The accumulation of the processed LHCP is enhanced by the addition to the chloroplasts of a precursor and a co-factor for chlorophyll biosynthesis. Using a model for the arrangement of the mature polypeptide in the thylakoid membrane as a guide, we have created mutations that lie within the mature coding region. We have studied the processing, the integration into thylakoid membranes, and the assembly into light-harvesting complexes of six of these deletions. Four different mutant LHCPs are found as processed proteins in the thylakoid membrane, but only one appears to have an orientation in the membrane that is similar to that of the wild type. No mutant LHCP appears in LHC II. The other two mutant LHCPs cannot be detected within the chloroplasts. We conclude that stable complex formation is not required for the processing and insertion of altered LHCPs into the thylakoid membrane. We discuss the results in light of our model.
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002734:Chlorophyll / Q000235:genetics / Q000502:physiology*; D002736:Chloroplasts / Q000378:metabolism* / Q000648:ultrastructure; D003001:Cloning, Molecular; D007425:Intracellular Membranes / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D010940:Plant Proteins / Q000235:genetics / Q000502:physiology*; D010944:Plants / Q000235:genetics; D011487:Protein Conformation; D011498:Protein Precursors / Q000096:biosynthesis; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism; D014357:Trypsin
D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D002734:Chlorophyll; D014357:Trypsin