Abstract
N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and its derivatives are putative calmodulin antagonists that bind to calmodulin and inhibit Ca2+/calmodulin-regulated enzyme activities. Autoradiographic studies using tritiated W-7 showed that this compound penetrates the cell membrane, is distributed mainly in the cytoplasm, and inhibits proliferation of Chinese hamster ovary K1 (CHO-K1) cells. Cytoplasmic [3H]W-7 was excluded completely within 6 hr after removal of [3H]W-7 from the culture medium. N-(6-aminohexyl)-1-naphthalenesulfonamide, an analogue of W-7 that interacts only weakly with calmodulin, proved to be a much weaker inhibitor of cell proliferation. CHO-K1 cells were synchronized by shaking during mitosis and then released into the cell cycle in the presence of 25 microM W-7 or 2.5 mM thymidine for 12 hr. Cell division was observed approximately 6 hr later. The results suggest that the effect of W-7 on cell proliferation might be through selective inhibition of the G1/S boundary phase, which is similar to the effect of excess thymidine. This pharmacological demonstration that cytoplasmic calmodulin is involved in cell proliferation is significant; W-7 and its derivatives may be useful tools for research on calmodulin and cell biology-related studies.
Full text
PDF![4354](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eae4/319788/620ecf354d9d/pnas00658-0395.png)
![4355](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eae4/319788/f12511122c0e/pnas00658-0396.png)
![4356](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eae4/319788/839e22384529/pnas00658-0397.png)
![4357](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eae4/319788/2bbdb5f15391/pnas00658-0398.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allan D., Michell R. H. A comparison of the effects of phytohaemagglutinin and of calcium ionophore A23187 on the metabolism of glycerolipids in small lymphocytes. Biochem J. 1977 May 15;164(2):389–397. doi: 10.1042/bj1640389. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boynton A. L., Whitfield J. F., MacManus J. P. Calmodulin stimulates DNA synthesis by rat liver cells. Biochem Biophys Res Commun. 1980 Jul 31;95(2):745–749. doi: 10.1016/0006-291x(80)90849-9. [DOI] [PubMed] [Google Scholar]
- Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase. Demonstration of an activator. Biochem Biophys Res Commun. 1970 Feb 6;38(3):533–538. doi: 10.1016/0006-291x(70)90747-3. [DOI] [PubMed] [Google Scholar]
- Dedman J. R., Welsh M. J., Means A. R. Ca2+-dependent regulator. Production and characterization of a monospecific antibody. J Biol Chem. 1978 Oct 25;253(20):7515–7521. [PubMed] [Google Scholar]
- Douglas W. W. Stimulus-secretion coupling: the concept and clues from chromaffin and other cells. Br J Pharmacol. 1968 Nov;34(3):451–474. doi: 10.1111/j.1476-5381.1968.tb08474.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HUMMEL J. P., DREYER W. J. Measurement of protein-binding phenomena by gel filtration. Biochim Biophys Acta. 1962 Oct 8;63:530–532. doi: 10.1016/0006-3002(62)90124-5. [DOI] [PubMed] [Google Scholar]
- Hidaka H., Asano M., Iwadare S., Matsumoto I., Totsuka T., Aoki N. A novel vascular relaxing agent, N-(6--aminohexyl)-5-chloro-1-naphthalensulfonamide which affects vascular smooth muscle actomyosin. J Pharmacol Exp Ther. 1978 Oct;207(1):8–15. [PubMed] [Google Scholar]
- Hidaka H., Asano T. Platelet cyclic 3':5'-nucleotide phosphodiesterase released by thrombin and calcium ionophore. J Biol Chem. 1976 Dec 10;251(23):7508–7516. [PubMed] [Google Scholar]
- Hidaka H., Naka M., Yamaki T. Effect of novel specific myosin light chain kinase inhibitors on Ca2+-activated Mg2+-ATPase of chicken gizzard actomyosin. Biochem Biophys Res Commun. 1979 Oct 12;90(3):694–699. doi: 10.1016/0006-291x(79)91883-7. [DOI] [PubMed] [Google Scholar]
- Hidaka H., Yamaki T., Asano M., Totsuka T. Involvement of calcium in cyclic nucleotide metabolism in human vascular smooth muscle. Blood Vessels. 1978;15(1-3):55–64. doi: 10.1159/000158153. [DOI] [PubMed] [Google Scholar]
- Hidaka H., Yamaki T., Naka M., Tanaka T., Hayashi H., Kobayashi R. Calcium-regulated modulator protein interacting agents inhibit smooth muscle calcium-stimulated protein kinase and ATPase. Mol Pharmacol. 1980 Jan;17(1):66–72. [PubMed] [Google Scholar]
- Hidaka H., Yamaki T., Totsuka T., Asano M. Selective inhibitors of Ca2+-binding modulator of phosphodiesterase produce vascular relaxation and inhibit actin-myosin interaction. Mol Pharmacol. 1979 Jan;15(1):49–59. [PubMed] [Google Scholar]
- Hidaka H., Yamaki T., Yamabe H. Two forms of Ca2+-dependent cyclic 3':5'-nucleotide phosphodiesterase from human aorta and effect of free fatty acids. Arch Biochem Biophys. 1978 Apr 30;187(2):315–321. doi: 10.1016/0003-9861(78)90040-1. [DOI] [PubMed] [Google Scholar]
- Kobayashi R., Tawata M., Hidaka H. Ca2+ regulated modulator protein interacting agents: inhibition of Ca2+-Mg2+-ATPase of human erythrocyte ghost. Biochem Biophys Res Commun. 1979 Jun 13;88(3):1037–1045. doi: 10.1016/0006-291x(79)91513-4. [DOI] [PubMed] [Google Scholar]
- Lestourgeon W. M., Forer A., Yang Y. Z., Bertram J. S., Pusch H. P. Contractile proteins. Major components of nuclear and chromosome non-histone proteins. Biochim Biophys Acta. 1975 Feb 27;379(2):529–552. [PubMed] [Google Scholar]
- MacManus J. P., Boynton A. L., Whitfield J. F. Cyclic AMP and calcium as intracycle regulators in the control of cell proliferation. Adv Cyclic Nucleotide Res. 1978;9:485–491. [PubMed] [Google Scholar]
- MacManus J. P., Whitfield J. F., Boynton A. L., Rixon R. H. Role of cyclic nucleotides and calcium in the positive control of cell proliferation. Adv Cyclic Nucleotide Res. 1975;5:719–734. [PubMed] [Google Scholar]
- Nagata T., Nawa T. A modification of dry-mounting technique for radioautography of water-soluble compounds. Histochemie. 1966;7(4):370–371. doi: 10.1007/BF00306625. [DOI] [PubMed] [Google Scholar]
- Seeman P., Staiman A., Chau-Wong M. The nerve impulse-blocking actions of tranquilizers and the binding of neuroleptics to synaptosome membranes. J Pharmacol Exp Ther. 1974 Jul;190(1):123–130. [PubMed] [Google Scholar]
- TERASIMA T., TOLMACH L. J. Growth and nucleic acid synthesis in synchronously dividing populations of HeLa cells. Exp Cell Res. 1963 Apr;30:344–362. doi: 10.1016/0014-4827(63)90306-9. [DOI] [PubMed] [Google Scholar]
- Tanaka T., Hidaka H. Hydrophobic regions function in calmodulin-enzyme(s) interactions. J Biol Chem. 1980 Dec 10;255(23):11078–11080. [PubMed] [Google Scholar]
- Tanaka T., Naka M., Hidaka H. Activation of myosin light chain kinase by trypsin. Biochem Biophys Res Commun. 1980 Jan 15;92(1):313–318. doi: 10.1016/0006-291x(80)91554-5. [DOI] [PubMed] [Google Scholar]
- Welsh M. J., Dedman J. R., Brinkley B. R., Means A. R. Calcium-dependent regulator protein: localization in mitotic apparatus of eukaryotic cells. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1867–1871. doi: 10.1073/pnas.75.4.1867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welsh M. J., Dedman J. R., Brinkley B. R., Means A. R. Tubulin and calmodulin. Effects of microtubule and microfilament inhibitors on localization in the mitotic apparatus. J Cell Biol. 1979 Jun;81(3):624–634. doi: 10.1083/jcb.81.3.624. [DOI] [PMC free article] [PubMed] [Google Scholar]
- XEROS N. Deoxyriboside control and synchronization of mitosis. Nature. 1962 May 19;194:682–683. doi: 10.1038/194682a0. [DOI] [PubMed] [Google Scholar]