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9 Use of a leucine clamp to demonstrate that IGF-I actively stimulates protein synthesis in normal humans. Physiology.org

In the first part of the reaction, formation of an electrophilic acyl-phosphate intermediate occurs by transfer of the -phosphate of ATP to -glutamylcysteine

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Extracted samples were vortexed at 4 C for 10 min, cleared by centrifugation (17,000 g , 10 min, 4 C) and then supernatants were analysed by LCMS