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Cell-free synthesis and processing of a large precursor of glutamate dehydrogenase of rat liver
Authors:Vicente Miralles  Vicente Felipo  Jose Hernández-Yago  Santiago Grisolía
Institution:Instituto de Investigaciones Citológicas de la Caja de Ahorros de Valencia Amadeo de Saboya 4, Valencia, Spain
Abstract:The mitochondrial matrix protein glutamate dehydrogenase of rat liver was synthesized in a cell-free reticulocyte lysate using mRNA from free or membrane-bound polysomes from rat liver. Immunoprecipitation of the (35S)methionine labeled translation mixture was performed using rabbit anti-glutamate dehydrogenase serum. Analysis after electrophoresis of the immunoprecipitate by fluorography of a dried sodium dodecyl sulfate/polyacrylamide gel showed that the glutamate dehydrogenase is synthesized ‘in vitro’ as a large precursor. A mitochondrial extract from rat liver processed the precursor synthesized “in vitro” to the mature form.
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