Induction of apoptosis by electrotransfer of positively charged proteins as Cytochrome C and Histone H1 into cells.
Cytochrome C (Cyt. C) is a mitochondrial protein inducing apoptosis when it is accumulated in the cytosol by a currently unknown mechanism, but regulated by the bcl-2 family of proteins. The linker Histone H1 is another basic protein with highly conservative structure, composition, and equal molecular weight, not changed during the evolution. An attempt was made to understand better the apoptotic processes by electroloading of leukemic cells, such as K562, HL-60, and SKW3, and human lymphocytes with positively charged proteins, such as Cyt. C, Histone H1, and methylated BSA albumin (mBSA). The triggering apoptotic processes followed by MTT test, FACS analysis, and DNA fragmentation after the electrotransfer of these proteins into the cells were observed. Histone H1 and mBSA induce the release of Cyt. C from rat liver mitochondria. Cytochrome C release was higher when mitochondria were in "high-energy" state. It is supposed that release of Cyt. C from mitochondria is due to the mechanical rupture of the outer mitochondrial membrane, rich in negatively charged groups, predominately due to cardiolipin. The reason for the morphological rupture of the outer mitochondial membrane could be the rigidification and segregation of the membrane and the destroyed membrane asymmetries of both monolayers in the presence of positively charged proteins at higher linear charges such as Histone H1. We suggested that Histone H1, at a given moment of activated signaling for apoptosis, could be not transported to the nucleus and could lead to the release of Cyt. C from the mitochondria in the cytoplasm. It is temping to speculate that Histone H1 has other physiological extranuclear functions involved in apoptosis.
Tsoneva
I
I
Institute of Biophysics, Bulg. Acad. Sci., Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria. itsoneva@obzor.bio21.bas.bg
Nikolova
B
B
Georgieva
M
M
Guenova
M
M
Tomov
T
T
Rols
M-P
MP
Berger
M R
MR
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000494:pharmacology*; D053938:DNA Fragmentation; D018274:Electroporation; D006657:Histones / Q000494:pharmacology*; D006801:Humans; D020014:K562 Cells; D008930:Mitochondria, Liver / Q000201:enzymology; D051381:Rats; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D006657:Histones; D012710:Serum Albumin, Bovine; D045304:Cytochromes c