Characterization of an initiating cell-free protein synthesis system derived from rabbit brain.

Abstract:

:Protein synthesis in the brain is known to be affected by a wide range of treatments. The detailed analysis of the mechanisms that are involved would be facilitated by the development of cell-free translation systems derived from brain tissue. To date, brain cell-free systems have not been fully characterized to demonstrate a capacity for initiation of translation. The following criteria were utilized to demonstrate that a cell-free protein synthesis system derived from rabbit brain was capable of initiation in vitro: (a) sensitivity of cell-free translation to the initiation inhibitor aurintricarboxylic acid (ATA); (b) binding of [35S]Met-tRNAf to 40S and 80S initiation complexes; (c) incorporation of labeled initiation methionine into high-molecular-weight proteins; and (d) the association of labeled exogenous mRNA with polysomes. The optimum conditions for amino acid incorporation in this system were 4 mM-Mg2+, 140 mM-K+, and pH 7.55. Incorporation was dependent on the addition of ATP, GTP, and an energy-generating system. Cell-free protein synthesis reflected the normal process, since a similar spectrum of proteins was synthesized in vitro and in vivo. This initiating cell-free translation system should have wide application in the analysis of the mechanisms whereby various treatments affect protein synthesis in the brain.

journal_name

J Neurochem

authors

Cosgrove JW,Brown IR

doi

10.1111/j.1471-4159.1981.tb01696.x

subject

Has Abstract

pub_date

1981-03-01 00:00:00

pages

1026-36

issue

3

eissn

0022-3042

issn

1471-4159

journal_volume

36

pub_type

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