In vivo measurement of synthesis rate of multiple plasma proteins in humans.
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.
Jaleel
Abdul
A
Division of Endocrinology, Diabetes, Metabolism, and Nutrition, Mayo Clinic, 200 First St. SW, 5-194 Joseph, Rochester, MN 55905, USA.
Nehra
Vandana
V
Persson
Xuan-Mai T
XM
Boirie
Yves
Y
Bigelow
Maureen
M
Nair
K Sreekumaran
KS
D000328:Adult; D001798:Blood Proteins / Q000032:analysis / Q000096:biosynthesis*; D002247:Carbon Isotopes; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D007553:Isotope Labeling; D008297:Male; D008894:Milk Proteins / Q000378:metabolism*; D014176:Protein Biosynthesis; D000067816:Whey Proteins
D001798:Blood Proteins; D002247:Carbon Isotopes; D008894:Milk Proteins; D000067816:Whey Proteins