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Stable isotope labeling tandem mass spectrometry (SILT) to quantify protein production and clearance rates

  • Randall J. BatemanEmail author
  • Ling Y. Munsell
  • Xianghong Chen
  • David M. Holtzman
  • Kevin E. Yarasheski
Articles

Abstract

In all biological systems, protein amount is a function of the rate of production and clearance. The speed of a response to a disturbance in protein homeostasis is determined by turnover rate. Quantifying alterations in protein synthesis and clearance rates is vital to understanding disease pathogenesis (e.g., aging, inflammation). No methods currently exist for quantifying production and clearance rates of low-abundance (femtomole) proteins in vivo. We describe a novel, mass spectrometry—based method for quantitating low-abundance protein synthesis and clearance rates in vitro and in vivo in animals and humans. The utility of this method is demonstrated with amyloid-β (Aβ), an important low-abundance protein involved in Alzheimer’s disease pathogenesis. We used in vivo stable isotope labeling, immunoprecipitation of Aβ from cerebrospinal fluid, and quantitative liquid chromatography electrospray-ionization tandem mass spectrometry (LC-ESI-tandem MS) to quantify human Aβ protein production and clearance rates. The method is sensitive and specific for stable isotope-labeled amino acid incorporation into CNS Aβ (+1% accuracy). This in vivo method can be used to identify pathophysiologic changes in protein metabolism and may serve as a biomarker for monitoring disease risk, progression, or response to novel therapeutic agents. The technique is adaptable to other macromolecules, such as carbohydrates or lipids.

Keywords

Clearance Rate Fractional Synthesis Rate Fractional Clearance Rate Mass Isotopomer Distribution Analysis Human Neuroglioma Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© American Society for Mass Spectrometry 2007

Authors and Affiliations

  • Randall J. Bateman
    • 1
    • 4
    • 5
    Email author
  • Ling Y. Munsell
    • 2
  • Xianghong Chen
    • 2
  • David M. Holtzman
    • 1
    • 3
    • 4
    • 5
  • Kevin E. Yarasheski
    • 2
  1. 1.Department of NeurologyWashington University School of MedicineSt. Louis
  2. 2.Department of MedicineWashington University School of MedicineSt. LouisUSA
  3. 3.Department of Molecular Biology and PharmacologyWashington University School of MedicineSt. LouisUSA
  4. 4.Hope Center for Neurological DisordersWashington University School of MedicineSt. LouisUSA
  5. 5.Alzheimer’s Disease Research CenterWashington University School of MedicineSt. LouisUSA

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