Abstract
The aim of this study was to analyze the N-terminal post-translational incorporation of arginine into cytosolic proteins from cultured cells and the in vitro incorporation of arginine into soluble proteins of PC12 cells after serum deprivation. Arginine incorporation was measured in the presence of protein synthesis inhibitors. None of the inhibitors used affected significantly the arginylation reaction while the novo synthesis of protein was reduced by 98%. Under these conditions, we found that of the total [14C]arginine incorporated into the proteins, around 20% to 40% was incorporated into the N-terminal position of soluble proteins by a post-translational mechanism. These results suggest that this post-translational aminoacylation may be a widespread reaction in neuronal and non-neuronal cells. We also found that in PC12 cells, the in vitro post-translational arginylation was 60% higher in apoptotic cells with respect to control cells. These findings suggest that the post-translational arginylation of proteins may be involved in programmed cell death.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Kaji, H., Novelli, G. D., and Kaji, A. A. 1963. A soluble amino acid incorporating system from rat liver. Biochim. Biophys. Acta. 76:474-477.
Barra, H. S., Rodriguez, J. A., Arce, C. A., and Caputto, R. 1973. A soluble preparation from rat brain that incorporates into its own proteins [14C]-arginine by a RNAse-sensitive system and [14C]-tyrosine by a RNAse insensitive system. J. Neurochem. 20:97-108.
Soffer, R. L. 1971. Enzymatic modification of proteins. Protein acceptor specificity in the arginine-transfer reaction. J. Biol. Chem. 246:1602-1606.
Shyne-Athwal, S., Riccio, R. V., Chakraborty, G., and Ingoglia, N. A. 1986. Protein modification by amino acid addition is increased in crushed sciatic but not optic nerves. Science. 231:603-605.
Soffer, R. L. 1973. Peptide acceptors in the arginine transfer reaction. J. Biol. Chem. 248:2918-2921.
Gower, D. J., and Tytell, M. 1986. Protein modification by RNA-dependent post-translational aminoacylation in synaptoplasm. J. Neurochem. 47:389-395.
Bongiovanni, G., Barra, H. S., and Hallak, M. E. 1994. Some common properties between a brain protein that is modified by a post-translational arginylation and the STOP protein. J. Neurochem. 63:2295-2299.
Hallak, M. E., and Bongiovanni, G. 1997. Post-translational arginylation of brain proteins. Neurochem. Res. 22:467-473.
Bosc, C., Cronk, J. D., Pirollet, F., Watterson, D. M., Haiech, J, Job D., and Margolis, R. 1996. Cloning, expression, and properties of microtubule-stabilizing protein STOP. Proc. Natl. Acad. Sci. USA, 93:2125-2130.
Guillaud, L., Bosc, C., Fourest-Lieuvin, A., Denarier, E., Pirollet, F., Lafanechè re, L., and Job, D. 1998. STOP proteins are responsible for the high degree of microtubule stabilization observed in neuronal cells. J. Cell Biol. 142:167-179.
Zanakis, M. F., Chakraborty, G., Sturman, J. A., and Ingoglia, N. A. 1984. Post-translational protein modification by amino acid addition in intact and regenerating axons of the rat sciatic nerve. J. Neurochem. 3:1286-1294.
Xu, N-S., Chakraborty, G., Hassankhani, A., and Ingoglia, N. A. 1993. N-terminal arginylation of proteins in explants of injured sciatic nerves and embryonic brain of rats. Neurochem. Res. 18:1117-1123.
Jones, M. E., Haire, M. F., Kloetzel, P. M., Mykles, D. L., and Schwartz, L. M. 1995. Changes in the structure and function of the Multicatalitic Proteinase (Proteosome) during programmed cell death in the intersegmental muscles of the hawkmoth, Manduca sexta. Dev. Biol. 169:436-447.
Bongiovanni, G., Fissolo, S., Barra, H. S., and Hallak, M. E. 1999. Post-translational arginylation of soluble rat brain proteins after whole body hypertermia. J. Neurosci. Res. 56:85-92.
Ferber, S., and Ciechanover, A. 1987. Role of arginine-tRNA in protein degradation by ubiquitin pathway. Nature 326:808-811.
Elias, S., and Ciechanover, A. 1990. Post-translational addition of arginine moiety to acidic NH2 termini of proteins is required for their recognition by ubiquitin-protein ligase. J. Biol. Chem. 265:15511-15517.
Bachmair, A., Finley, D., and Varshavsky, A. 1986. In vivo half-life of a protein is a function of its amino-terminal residue. Science. 234:179-186.
Patel, T., Gores, G. J., and Kaufmann, S. H. 1996. The role of proteases during apoptosis. Faseb J. 10:587-597.
Schawrtz, L., Smith, S., Jones, M., and Osborne, B. 1993. Do all programmed cell deaths occur via apoptosis?. Proc. Natl. Acad. Sci. USA. 90:980-984.
Greene, L. A., and Tischler, A. S. 1976. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to NGF. Proc. Natl. Acad. Sci. USA. 73:2424-2428.
Pettman, J. C. L., and Sensenbrenner, M. 1979. Morphological and biochemical maturation of neurons cultured in the absence of glial cells. Nature. 281:378-380.
Hallak, M. E., Bongiovanni, G., and Barra, H. S. 1991. The post-translational arginylation of protein in different regions of the rat brain. J. Neurochem. 57:1735-1739.
Levy, A. L., 1955. Recent developments in techniques for terminal and sequence studies in peptides and proteins. Methods of Biochemical Analysis (Edited by Glick D.) 2:360. Interscience, New York.
Edman, P. 1950. Method for determination of the amino acid sequence in peptides. Acta Chem. Scand. 4:283-293.
De Lange, R. J., Fambrough, D., Smith, E., and Bonner, J. 1968. Calf and pea histone IV. J. Biol. Chem. 243:5906-5913.
Gavrieli, Y., Sherman, Y., and Ben-Sasson, S. A. 1992. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell. Biol. 119:493-501.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265-275.
Rukenstein, A., Rydel, R. E., and Greene, L. A. 1991. Multiple agents rescue PC12 cells from serum-free cell death by translation and transcription independent mechanisms. J. Neurosci. 11:2552-2563.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fissolo, S., Bongiovanni, G., Decca, M.B. et al. Post-Translational Arginylation of Proteins in Cultured Cells. Neurochem Res 25, 71–76 (2000). https://doi.org/10.1023/A:1007539532469
Issue Date:
DOI: https://doi.org/10.1023/A:1007539532469