Abstract
Rhodanese (thiosulfate cyanide sulfurtransferase; E.C. 2.8.1.1) is a mitochondrial enzyme that is unprocessed after import. We describein vitro experiments showing that partially folded rhodanese can interact with lipid bilayers. The interaction was monitored by measuring the ability of rhodanese to disrupt small unilamellar vesicles composed of phosphatidylserine and to release 6-carboxyfluorescein that was trapped in the liposomes. Partially folded rhodanese, derived by dilution of urea-unfolded enzyme, efficiently induced liposome leakage. Native rhodanese had no effect on liposome integrity. Liposome disruption progressively decreased as rhodanese was given the opportunity to refold or aggregate before introduction of the liposomes. A synthetic 23 amino acid peptide representing the N-terminal sequence of rhodanese was very efficient at disrupting the liposomes. Shorter peptides chosen from within this sequence (residues 11–23 or residues 1–17) had no effect on liposome disruption. A peptide representing the tether region that connects the domains of the enzyme was also without effect. These results are consistent with the hypothesis that the N-terminal sequence of rhodanese is an uncleaved leader sequence, and can interact with membrane components that are involved in the mitochondrial uptake of this protein.
Similar content being viewed by others
References
Allison, D. S., and Schatz, G. (1986).Proc. Nat. Acad. Sci. 83, 9011–9015.
Aoyagi, H., Lee, S., Nakamura, H., Park, N. G., and Kato, T. (1988).Int. J. Pep. Prot. Res. 32, 406–414.
Barenholz, Y., Gibbes, D., Litman, B. J., Goll, J., Thompson, T. E., and Carlson, F. D. (1977).Biochemistry 16, 2806–2810.
Bartlett, G. R. (1959).J. Biol. Chem. 234, 466–468.
Endo, T., and Schatz, G. (1988).EMBO J. 7, 1153–1158.
Gierasch, L. M. (1989).Biochemistry 28, 923–930.
Grant, E., Jr. (1991). Ph.D. dissertation, Uniformed Services of the Health Sciences, Bethesda, Maryland.
Hol, W. G. J., Lijk, L. J., and Kalk, K. H. (1983).Fundam. Appl. Toxicol. 3, 370–376.
Horwich, A. L., Cheng, M., West, A., and Pollock, R. A. (1991).Current Topics Micro. Immunol. 170, 1–42.
Horowitz, P. (1978).Anal. Biochem. 86, 751–753.
Kurzban, G. P., and Horowitz, P. M. (1991).Protein. Expression Purif. 2, 379–384.
Landry, S. J., and Gierasch, L. M. (1991).Biochemistry 30, 7359–7362.
Lee, S., Mihara, H., Aoyagi, H., Kato, T., Izumiya, N., and Yamasaki, N. (1986).Biochim. Biophys. Acta 862, 211–219.
Lemire, B. D., Fankhauser, C., Baker, A., and Schatz, G. (1989).J. Biol. Chem. 264, 20,206–20,215.
Mendoza, J. A., Rogers, E., Lorimer, G. H., and Horowitz, P. M. (1991).J. Biol. Chem. 266, 13,587–13,591.
Merrill, G. A., Miller, D., Chirgwin, J., and Horowitz, P. M. (1992).J. Prot. Chem. (in press).
Miller, D. M., Delgado, R., Chirgwin, J. M., Hardies, S. C., and Horowitz, P. M. (1991).J. Biol. Chem. 266, 4686–4691.
Miller, D. M., Kurzban, G. P., Mendoza, J. A., Chirgwin, J. M., Hardies, S. C., and Horowitz, P. M. (1992).Biochem. Biophys. Acta (in press).
Ou, W.-J., Ito, A., Umeda, M., Inoue, K., and Omura, T. (1988).J. Biochem. 103, 589–595.
Pak, Y. K., and Weiner, H. (1990).J. Biol. Chem. 265, 14,298–14,307.
Pfanner, N., and Neupert, W. (1989).Curr. Opin. Cell Biol. 1, 624–629.
Ploegman, J. H., Drent, G. H., Kalk, K. H., Hol, W. G. J., Heinrikson, R. L., Keim, P., Weng, L., and Russell, J. (1978).Nature 273, 1245–1249.
Roise, D., Horvath, S. J., Tomich, J. M., Richards, J. H., and Schatz, G. (1986).EMBO J. 5, 1327–1334.
Schatz, G. (1989). Robert A. Welch Foundation Conference on Chem. Res. XXXIII Membrane Proteins: Targeting and Transduction, Chapter 4, pp. 77–95.
Scherer, P. E., Krieg, U. C., Hwang, S. T., Vestweber, D., and Schatz, G. (1990).EMBO J. 9, 4315–4322.
Sorbo, B. (1953).Acta Chem. Scand. 7, 1129–1136.
Verner, K., and Schatz, G. (1987).EMBO J. 6, 2449–2456.
Weinhues, U., Becker, K., Schleyer, M., Guiard, B., Tropschug, M., Horwich, A. L., Pfanner, N., and Neupert, W. (1991).J. Cell. Biol. 115, 1601–1609.
Westley, J. (1973).Adv. Enzymol. Relat. Areas Mol. Biol. 39, 327–368.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mendoza, J.A., Grant, E. & Horowitz, P.M. Partially folded rhodanese or its N-terminal sequence can disrupt phospholipid vesicles. J Protein Chem 12, 65–69 (1993). https://doi.org/10.1007/BF01024916
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01024916