Visualization of antibody binding to the photosynthetic membrane: The transmembrane orientation of cytochrome b-559

摘要: We have used immuno-gold labeling and electron microscopy to studythe topography of thylakoid membrane polypeptides. Thylakoid vesicles formed by passage through a French press were adsorbed onto plastic film supported an microscope grid processed for single or double labeling. Aftershadowing with platinum, the inside-out right-side-out identified their distinctive morphologies. Right-side-out labeled monoclonal antibody recognizing epitope located in trypsin-cleaved, N- terminal portion LHC II apoprotein, CF 1 . A α-subunit cytochrome b-559 reacted synthetic tridecapeptide corresponding C-terminal polypeptide. Both this apolyclonal peptide exclusively, indicating that polypeptide C-terminus was exposed on lumenal (exoplasmic) surface membrane.