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Development and use of domain-specific antibodies in a characterization of the large subunits of soybean photosystem 1The 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.
Document ID
20040090217
Acquisition Source
Headquarters
Document Type
Reprint (Version printed in journal)
Authors
Henry, R. L.
(Kansas State University Manhattan, United States)
Takemoto, L. J.
Murphy, J.
Gallegos, G. L.
Guikema, J. A.
Spooner, B. S.
Date Acquired
August 21, 2013
Publication Date
January 1, 1992
Publication Information
Publication: Plant physiology and biochemistry : PPB / Societe francaise de physiologie vegetale
Volume: 30
Issue: 3
ISSN: 0981-9428
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: NAGW-2328
CONTRACT_GRANT: NAGW-1197
Distribution Limits
Public
Copyright
Other
Keywords
NASA Program NSCORT
NASA Discipline Cell Biology
NASA Discipline Number 93-10
Non-NASA Center
NASA Discipline Developmental Biology

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