In vivo measurement of synthesis rate of multiple plasma proteins in humans

Abdul Jaleel, Vandana Nehra, Xuan Mai T. Persson, Yves Boirie, Maureen Bigelow, K. Sreekumaran Nair

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Advances in quantitative proteomics have facilitated the measurement of large-scale protein quantification, which represents net changes in protein synthesis and breakdown. However, measuring the rate of protein synthesis is the only way to determine the translational rate of gene transcripts. Here, we report a technique to measure the rate of incorporation of amino acids from ingested protein labeled with stable isotope into individual plasma proteins. This approach involves three steps: 1) production of stable isotope-labeled milk whey protein, oral administration of this intrinsically labeled protein, and subsequent collection of blood samples; 2) fractionation of the plasma and separation of the individual plasma proteins by a combination of anion exchange high-pressure liquid chromatography and gel electrophoresis; and 3) identification of individual plasma proteins by tandem mass spectrometry and measurement of stable isotopic enrichment of these proteins by gas chromatography-mass spectrometry. This method allowed the measurement of the fractional synthesis rate (FSR) of 29 different plasma proteins by using the same precursor pool. We noted a 30-fold difference in FSR of different plasma proteins with a wide range of physiological functions. This approach offers a tremendous opportunity to study the regulation of plasma proteins in humans in many physiological and pathological states.

Original languageEnglish (US)
Pages (from-to)E190-E197
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume291
Issue number1
DOIs
StatePublished - 2006

Keywords

  • Milk proteins
  • Phenylalanine
  • Plasma protein synthesis
  • Proteomics
  • Stable isotope

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Physiology (medical)

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