Lipid modifications of G protein subunits: Myristoylation of G(oα) increases its affinity for βγ

M. E. Linder, Iok-Hou Pang, R. J. Duronio, J. I. Gordon, P. C. Sternweis, A. G. Gilman

Research output: Contribution to journalArticle

196 Citations (Scopus)

Abstract

Myristoylated recombinant proteins can be synthesized in Escherichia coli by concurrent expression of the enzyme myristoyl-CoA:protein N-myristoyl-transferase with its protein substrates (Duronio, R.J., Jackson-Machelski, E., Heuckeroth, R.O., Olins, P.O., Devine, C.S., Yonemoto, W., Slice, L.W., Taylor, S. S., and Gordon, J.I. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 1506-1510). Expression of the G protein subunit G(oα) in this system results in the synthesis of two forms of the protein; these were separated on a column of heptylamine-Sepharose. Purification of the more abundant form of G(oα) yielded a product that has a blocked amino terminus. Chemical analysis of the fatty acids released by acid hydrolysis of the protein revealed myristic acid. The second form of the protein was not myristoylated. Myristoylated and nonmyristoylated recombinant G(oα) were compared with brain G(oα) (which is myristoylated) for their ability to interact with G protein βγ subunits. The nonmyristoylated recombinant protein clearly had a reduced affinity for βγ, while the myristoylated recombinant protein was indistinguishable from native G(oα) in its subunit interactions. Thus, myristoylation increases the affinity of α subunits for βγ. We propose that the function of myristoylation of G protein α subunits is, at least in part, to facilitate formation of the heterotrimer and the localization of α to the plasma membrane.

Original languageEnglish
Pages (from-to)4654-4659
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number7
StatePublished - 18 Jul 1991

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Protein Subunits
GTP-Binding Proteins
Recombinant Proteins
Lipids
Proteins
Distillation columns
Myristic Acid
Cell membranes
Sepharose
Escherichia coli
Purification
Hydrolysis
Brain
Fatty Acids
Cell Membrane
Acids
Substrates
Enzymes
Chemical analysis

Cite this

Linder, M. E., Pang, I-H., Duronio, R. J., Gordon, J. I., Sternweis, P. C., & Gilman, A. G. (1991). Lipid modifications of G protein subunits: Myristoylation of G(oα) increases its affinity for βγ. Journal of Biological Chemistry, 266(7), 4654-4659.
Linder, M. E. ; Pang, Iok-Hou ; Duronio, R. J. ; Gordon, J. I. ; Sternweis, P. C. ; Gilman, A. G. / Lipid modifications of G protein subunits : Myristoylation of G(oα) increases its affinity for βγ. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 7. pp. 4654-4659.
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Linder, ME, Pang, I-H, Duronio, RJ, Gordon, JI, Sternweis, PC & Gilman, AG 1991, 'Lipid modifications of G protein subunits: Myristoylation of G(oα) increases its affinity for βγ', Journal of Biological Chemistry, vol. 266, no. 7, pp. 4654-4659.

Lipid modifications of G protein subunits : Myristoylation of G(oα) increases its affinity for βγ. / Linder, M. E.; Pang, Iok-Hou; Duronio, R. J.; Gordon, J. I.; Sternweis, P. C.; Gilman, A. G.

In: Journal of Biological Chemistry, Vol. 266, No. 7, 18.07.1991, p. 4654-4659.

Research output: Contribution to journalArticle

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T2 - Myristoylation of G(oα) increases its affinity for βγ

AU - Linder, M. E.

AU - Pang, Iok-Hou

AU - Duronio, R. J.

AU - Gordon, J. I.

AU - Sternweis, P. C.

AU - Gilman, A. G.

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N2 - Myristoylated recombinant proteins can be synthesized in Escherichia coli by concurrent expression of the enzyme myristoyl-CoA:protein N-myristoyl-transferase with its protein substrates (Duronio, R.J., Jackson-Machelski, E., Heuckeroth, R.O., Olins, P.O., Devine, C.S., Yonemoto, W., Slice, L.W., Taylor, S. S., and Gordon, J.I. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 1506-1510). Expression of the G protein subunit G(oα) in this system results in the synthesis of two forms of the protein; these were separated on a column of heptylamine-Sepharose. Purification of the more abundant form of G(oα) yielded a product that has a blocked amino terminus. Chemical analysis of the fatty acids released by acid hydrolysis of the protein revealed myristic acid. The second form of the protein was not myristoylated. Myristoylated and nonmyristoylated recombinant G(oα) were compared with brain G(oα) (which is myristoylated) for their ability to interact with G protein βγ subunits. The nonmyristoylated recombinant protein clearly had a reduced affinity for βγ, while the myristoylated recombinant protein was indistinguishable from native G(oα) in its subunit interactions. Thus, myristoylation increases the affinity of α subunits for βγ. We propose that the function of myristoylation of G protein α subunits is, at least in part, to facilitate formation of the heterotrimer and the localization of α to the plasma membrane.

AB - Myristoylated recombinant proteins can be synthesized in Escherichia coli by concurrent expression of the enzyme myristoyl-CoA:protein N-myristoyl-transferase with its protein substrates (Duronio, R.J., Jackson-Machelski, E., Heuckeroth, R.O., Olins, P.O., Devine, C.S., Yonemoto, W., Slice, L.W., Taylor, S. S., and Gordon, J.I. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 1506-1510). Expression of the G protein subunit G(oα) in this system results in the synthesis of two forms of the protein; these were separated on a column of heptylamine-Sepharose. Purification of the more abundant form of G(oα) yielded a product that has a blocked amino terminus. Chemical analysis of the fatty acids released by acid hydrolysis of the protein revealed myristic acid. The second form of the protein was not myristoylated. Myristoylated and nonmyristoylated recombinant G(oα) were compared with brain G(oα) (which is myristoylated) for their ability to interact with G protein βγ subunits. The nonmyristoylated recombinant protein clearly had a reduced affinity for βγ, while the myristoylated recombinant protein was indistinguishable from native G(oα) in its subunit interactions. Thus, myristoylation increases the affinity of α subunits for βγ. We propose that the function of myristoylation of G protein α subunits is, at least in part, to facilitate formation of the heterotrimer and the localization of α to the plasma membrane.

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