DOI: 10.1111/j.1471-4159.1980.tb11197.x. <i>In Vitro</i> Synthesis of the Myelin Basic Proteins: Subcellular Site of Synthesis

In Vitro Synthesis of the Myelin Basic Proteins: Subcellular Site of Synthesis

10.1111/j.1471-4159.1980.tb11197.x

Crossref journal-article

Wiley

Journal of Neurochemistry (311)

Abstract

Abstract: Brains of 3‐week‐old C57BL/6J mice were homogenized and fractionated into several subcellular components, each of which was examined for ability to synthesize the myelin basic proteins (MBPs) in vitro. Myelin basic proteins were purified from incubation mixtures by conventional means. That the products of synthesis were the myelin basic proteins was established by solubility at pH 3, co‐chromatography with authentic proteins on carboxymethylcellulose and co‐migration with standards in two different polyacrylamide gel electrophoretic systems. The fractions examined for their ability to synthesize MBPs were the whole homogenate, postnuclear supernatant, postmitochondrial supernatant, crude mitochondrial pellet, free ribosomes and bound ribosomes. Although there was no requirement for exogenous energy sources for protein synthesis in the whole homogenate, as the homogenate was fractionated an increasing requirement emerged. Most of the label in the MBP preparations from whole homogenate and postnuclear supernatant incubations migrated with the large (L) and small (S) MBPs on gel electrophoresis; however, as the homogenate was subfractionated and incubated, a greater percentage of the label migrated more slowly than L and S on acetic acid‐urea gels. To show synthesis of the MBPs the L and S bands were cut out of these gels and rerun on sodium dodecylsulfate gels. Alternatively, MBP preparations were subjected directly to two‐dimensional gel electrophoresis and the bands corresponding to L and S were excised and counted. With this method only the whole homogenate, postnuclear supernatant, postmitochondrial supernatant and free ribosomes were observed to synthesize the MBPs in vitro. The “bound” ribosomes were not observed to synthesize significant amounts of the MBPs, incubated either intact or released from the membrane. It was concluded that the free ribosomes are the principal site of synthesis of the myelin basic proteins in the brain.

Bibliography

Campagnoni, A. T., Carey, G. D., & Yu, Y. (1980). In Vitro Synthesis of the Myelin Basic Proteins: Subcellular Site of Synthesis. Journal of Neurochemistry, 34(3), 677â686. Portico.

Authors 3

Anthony T. Campagnoni (first)
Gregory D. Carey (additional)
Yie‐Teh Yu (additional)

References 27 Referenced 36

{'key': 'e_1_2_2_2_1', 'first-page': '87', 'article-title': 'Immunohistochemical localization of myelin proteolipid protein to oligodendrocytes in developing brain', 'volume': '10', 'author': 'Agrawal H. C.', 'year': '1979', 'journal-title': 'Trans. Am. Soc. Neurochem.'} / Trans. Am. Soc. Neurochem. / Immunohistochemical localization of myelin proteolipid protein to oligodendrocytes in developing brain by Agrawal H. C. (1979)
10.1073/pnas.74.8.3360
{'key': 'e_1_2_2_4_1', 'first-page': '183', 'volume-title': 'Mechanisms, Regulation and Special Functions of Protein Synthesis in the Brain', 'author': 'Benjamins J. A.', 'year': '1977'} / Mechanisms, Regulation and Special Functions of Protein Synthesis in the Brain by Benjamins J. A. (1977)
10.1007/BF00964057
10.1111/j.1471-4159.1977.tb10430.x
10.1016/0003-9861(78)90258-8
10.1111/j.1471-4159.1979.tb11709.x
10.1007/978-1-4757-4395-1_3
10.1073/pnas.74.3.1082
10.1016/0005-2787(65)90025-0
10.1042/bst0040742 / Biochem. Soc. Trans. / Developmental changes in the composition of rat brain myelin: Relationship between the two basic proteins by Giorgi P. P. (1976)
10.1073/pnas.70.5.1526
10.1042/bj1360825 / Biochem. J. / Preferential synthesis of a membrane‐associated protein by free polyribosomes by Lowe D. (1973)
10.1016/S0021-9258(19)52451-6
10.1016/0006-8993(77)90220-7
10.1016/0003-9861(61)90009-1
10.1016/0304-4157(76)90010-1
10.1016/0003-9861(69)90042-3
10.1111/j.1471-4159.1977.tb10422.x
10.1126/science.402030
{'key': 'e_1_2_2_22_1', 'first-page': '1784', 'article-title': 'In vitro incorporation of [3H]leucine into myelin basic protein', 'volume': '37', 'author': 'Shapira R.', 'year': '1978', 'journal-title': 'Fed. Proc.'} / Fed. Proc. / In vitro incorporation of [3H]leucine into myelin basic protein by Shapira R. (1978)
10.1111/j.1471-4159.1971.tb12004.x
10.1007/BF01217922
{'key': 'e_1_2_2_25_1', 'first-page': '86', 'article-title': 'Immunochemical demonstration of myelin‐associated glycoprotein', 'volume': '10', 'author': 'Sternberger N. H.', 'year': '1979', 'journal-title': 'Trans. Am. Soc. Neurochem.'} / Trans. Am. Soc. Neurochem. / Immunochemical demonstration of myelin‐associated glycoprotein by Sternberger N. H. (1979)
10.1016/S0021-9258(18)94333-4
10.1073/pnas.69.8.2326
10.1016/S0003-9861(73)80009-8

Dates

Type	When
Created	18 years, 10 months ago (Oct. 5, 2006, 6:15 a.m.)
Deposited	1 year, 9 months ago (Nov. 4, 2023, 4:42 a.m.)
Indexed	1 month, 3 weeks ago (July 7, 2025, 2:28 a.m.)
Issued	45 years, 5 months ago (March 1, 1980)
Published	45 years, 5 months ago (March 1, 1980)
Published Online	18 years, 10 months ago (Oct. 5, 2006)
Published Print	45 years, 5 months ago (March 1, 1980)

Funders 0

None

BibTeX

@article{Campagnoni_1980, title={In Vitro Synthesis of the Myelin Basic Proteins: Subcellular Site of Synthesis}, volume={34}, ISSN={1471-4159}, url={http://dx.doi.org/10.1111/j.1471-4159.1980.tb11197.x}, DOI={10.1111/j.1471-4159.1980.tb11197.x}, number={3}, journal={Journal of Neurochemistry}, publisher={Wiley}, author={Campagnoni, Anthony T. and Carey, Gregory D. and Yu, Yie‐Teh}, year={1980}, month=mar, pages={677–686} }

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