Incorporation of monoclonal antibodies in living rat pheochromocytoma PC12 cells. Evidence for the intracellular formation of immune complex between the incorporated antibody and a target protein.
PC12 cells permeabilized with a low concentration of digitonin (5 microM) under controlled conditions were loaded with monoclonal antibodies (MoAb) against the regulatory subunit type II (RII) of cAMP-dependent protein kinase. After digitonin removal from the nutrient medium (DMEM) the loaded cells repaired within 20-30 min and recontinued growth. The inserted MoAb stayed in the repaired cells at least for several hours. MoAb inhibiting the cAMP binding activity of neural RII [Grozdova et al. (1992) Biochem. Int. 27, 811-822; Sveshnikova et al. (1996) Biochem. Int. 39, 1063-1070] were shown to bind the target antigen inside the cells and influence the properties of intracellular protein kinases.
Alexandrova
N A
NA
Polymer Department, Chemical Faculty, Moscow State University, Russia.
Sveshnikov
P G
PG
Nagradova
N K
NK
Grozdova
I D
ID
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D000937:Antigen-Antibody Reactions; D002463:Cell Membrane Permeability / Q000187:drug effects; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases / Q000276:immunology / Q000378:metabolism; D004072:Digitonin / Q000494:pharmacology; D005057:Eukaryotic Cells / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D007202:Indicators and Reagents / Q000494:pharmacology; D016716:PC12 Cells / Q000276:immunology*; D011485:Protein Binding; D051381:Rats
D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases; D004072:Digitonin