English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Isolation and structural characterization of trimeric cyanobacterial photosystem I complex with the help of recombinant antibody fragments

MPS-Authors
/persons/resource/persons250817

Tsiotis,  Georgios
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;
Maurice E. Muller-Institute for Microscopical Structural Biology at the Biozentrum, University of Basel, Switzerland ;

/persons/resource/persons137691

Haase,  Winfried
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

/persons/resource/persons137800

Michel,  Hartmut
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

External Ressource
No external resources are shared
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)
There is no public supplementary material available
Citation

Tsiotis, G., Haase, W., Engel, A., & Michel, H. (1995). Isolation and structural characterization of trimeric cyanobacterial photosystem I complex with the help of recombinant antibody fragments. European Journal of Biochemistry, 231(3), 823-830. doi:10.1111/j.1432-1033.1995.tb20767.x.


Cite as: http://hdl.handle.net/21.11116/0000-0007-44D2-F
Abstract
A monoclonal antibody was derived from mice immunized with the native trimeric, photosystem I (PSI) complex from the cyanobacterium Synechococcus PCC 7002 which reacts with a conformational epitope of the PSI complex. As seen by immunoelectron microscopy, the mAb bound to the stromal side of the thylakoid membranes. The DNA sequence encoding variable regions of the mAb was cloned into recombinant plasmids, sequenced and expressed in Escherichia coli. ELISA, Western blots and immunoelectron microscopy provided evidence that the expressed paired variable domain (Fv) fragments bind to the antigen in the same way as the parent mAb. A one-step purification was applied to purify the trimeric PSI complex using an affinity tag attached to the Fv fragment. Analysis by gel electrophoresis and N-terminal sequencing revealed the presence of the psaA, psaB, psaC, psaD, psaE, psaF and psaL gene products. The antenna size of the isolated PSI/Fv was 139 +/- 9 chlorophyll a/primary electron donor. Flash-induced absorption-change measurements showed that the complex exhibited electron transfer from the primary electron donor, P700, to the Fe-S center, FA/FB. The position of the bound Fv fragment on the trimeric PSI surface was determined by high-resolution electron microscopy and digital image processing.