Detection of apoptosis using the C2A domain of synaptotagmin I.
Binding of annexin V or the C2A domain of synaptotagmin I to phosphatidylserine expressed on the surface of apoptotic cells can, when labeled with appropriate probe molecules, be used to detect the presence of apoptosis using radionuclide, magnetic resonance, and optical imaging techniques. The preparation of a biotinylated C2A-GST fusion protein is described, and its capability, when used in conjunction with fluorescein-labeled streptavidin, of detecting apoptotic cells by flow cytometry is compared directly with the performance of a commercial preparation of fluorescein-labeled annexin V. Biotinylated C2A-GST, when used in conjunction with streptavidin-conjugated superparamagnetic iron oxide nanoparticles or Gd-chelate-avidin conjugates, was shown to be capable of detecting apoptotic cells using T(2)-weighted or T(1)-weighted magnetic resonance imaging experiments, respectively.
Jung
Hyo-Il
HI
Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1GA, UK.
Kettunen
Mikko I
MI
Davletov
Bazbek
B
Brindle
Kevin M
KM
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002135:Calcium-Binding Proteins / Q000032:analysis*; D045744:Cell Line, Tumor; D003175:Complement C2 / Q000032:analysis*; D050678:Complement C2a; D008562:Membrane Glycoproteins / Q000032:analysis*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis*; D050863:Synaptotagmin I; D050857:Synaptotagmins
D002135:Calcium-Binding Proteins; D003175:Complement C2; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D050863:Synaptotagmin I; C494311:Syt1 protein, mouse; D050857:Synaptotagmins; D050678:Complement C2a