Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 47-52 |
| Number of pages | 6 |
| Journal | Analytical Biochemistry |
| Volume | 204 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1992 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Adenine and guanine nucleotide content of triton-extracted cytoskeletal fractions of nonmuscle cells. / Angelastro, James M; Purich, Daniel L.
In: Analytical Biochemistry, Vol. 204, No. 1, 1992, p. 47-52.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Adenine and guanine nucleotide content of triton-extracted cytoskeletal fractions of nonmuscle cells
AU - Angelastro, James M
AU - Purich, Daniel L.
PY - 1992
Y1 - 1992
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0026708034&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026708034&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(92)90137-V
DO - 10.1016/0003-2697(92)90137-V
M3 - Article
C2 - 1514695
AN - SCOPUS:0026708034
VL - 204
SP - 47
EP - 52
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -