Development and use of domain-specific antibodies in a characterization of the large subunits of soybean photosystem 1

The molecular architecture of the soybean photosystem 1 reaction center complex was examined using a combination of surface labeling and immunological methodology on isolated thylakoid membranes. Synthetic peptides (12 to 14 amino acids in length) were prepared which correspond to the N-terminal re...

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Veröffentlicht in:Plant physiology and biochemistry : PPB. - 1991. - 30(1992), 3 vom: 01., Seite 357-64
1. Verfasser: Henry, R L (VerfasserIn)
Weitere Verfasser: Takemoto, L J (BerichterstatterIn), Murphy, J, Gallegos, G L, Guikema, J A, Spooner, B S
Format: Aufsatz
Sprache:English
Veröffentlicht: 1992
Zugriff auf das übergeordnete Werk:Plant physiology and biochemistry : PPB
Schlagworte:Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. NASA Discipline Cell Biology NASA Discipline Developmental Biology NASA Discipline Number 93-10 NASA Program NSCORT Non-NASA Center Bacterial Proteins Immune Sera mehr... Light-Harvesting Protein Complexes Membrane Proteins PSI-A protein, Synechococcus Photosynthetic Reaction Center Complex Proteins Photosystem I Protein Complex Succinimides biotinyl N-hydroxysuccinimide ester photosystem I, psaB subunit Biotin 6SO6U10H04 Cyanogen Bromide OS382OHJ8P
Beschreibung
Zusammenfassung:The molecular architecture of the soybean photosystem 1 reaction center complex was examined using a combination of surface labeling and immunological methodology on isolated thylakoid membranes. Synthetic peptides (12 to 14 amino acids in length) were prepared which correspond to the N-terminal regions of the 83 and 82.4 kDa subunits of photosystem 1 (the PsaA and PsaB proteins, respectively). Similarly, a synthetic peptide was prepared corresponding to the C-terminal region of the PsaB subunit. These peptides were conjugated to a carrier protein, and were used for the production of polyclonal antibodies in rabbits. The resulting sera could distinguish between the PsaA and PsaB photosystem 1 subunits by Western blot analysis, and could identify appropriate size classes of cyanogen bromide cleavage fragments as predicted from the primary sequences of these two subunits. When soybean thylakoid membranes were surface-labeled with N-hydroxysuccinimidobiotin, several subunits of the complete photosystem 1 lipid/protein complex incorporated label. These included the light harvesting chlorophyll proteins of photosystem 1, and peptides thought to aid in the docking of ferredoxin to the complex during photosynthetic electron transport. However, the PsaA and PsaB subunits showed very little biotinylation. When these subunits were examined for the domains to which biotin did attach, most of the observed label was associated with the N-terminal domain of the PsaA subunit, as identified using a domain-specific polyclonal antisera
Beschreibung:Date Completed 14.06.1995
Date Revised 13.12.2023
published: Print
Citation Status MEDLINE
ISSN:1873-2690