Adenine and guanine nucleotide content of Triton-extracted cytoskeletal fractions of nonmuscle cells.
Determination of the adenine and guanine nucleotides in Triton X-100-extracted cytoskeletal fractions was utilized to estimate the actin and tubulin content of the assembled cytoskeletons in nonmuscle cells. Results with stable cell lines (i.e., rat pheochromocytoma PC12 and neuroblastoma NB41A3) and with primary cultures (i.e., human foreskin fibroblasts and chick embryonic dorsal root ganglion neurons) exhibited levels of cytoskeletal fraction ADP and GDP consistent with their assembly-induced nucleoside-5'-triphosphatase activities only previously analyzed in vitro. Likewise, estimates of actin and tubulin content fall in the range of values obtained by other experimental approaches. In contrast, analysis of whole cell nucleotides showed high [ATP]/[ADP] and [GTP]/[GDP] ratios, suggesting there is little, if any, contamination of the cytoskeletal nucleotide pool by other cellular nucleotides.
Angelastro
J M
JM
Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville 32610-0245.
Purich
D L
DL
D000227:Adenine Nucleotides / Q000032:analysis*; D000244:Adenosine Diphosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003599:Cytoskeleton / Q000737:chemistry*; D006150:Guanine Nucleotides / Q000032:analysis*; D006153:Guanosine Diphosphate / Q000032:analysis; D006160:Guanosine Triphosphate / Q000032:analysis; D006801:Humans; D017830:Octoxynol; D011092:Polyethylene Glycols; D014407:Tumor Cells, Cultured / Q000737:chemistry
D000227:Adenine Nucleotides; D006150:Guanine Nucleotides; D006153:Guanosine Diphosphate; D011092:Polyethylene Glycols; D000244:Adenosine Diphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D017830:Octoxynol