Format

Send to

Choose Destination
J Am Soc Mass Spectrom. 2007 Jun;18(6):997-1006. Epub 2007 Feb 22.

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

Author information

1
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri 63110, USA. batemanr@wustl.edu

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-beta (Abeta), an important low-abundance protein involved in Alzheimer's disease pathogenesis. We used in vivo stable isotope labeling, immunoprecipitation of Abeta from cerebrospinal fluid, and quantitative liquid chromatography electrospray-ionization tandem mass spectrometry (LC-ESI-tandem MS) to quantify human Abeta protein production and clearance rates. The method is sensitive and specific for stable isotope-labeled amino acid incorporation into CNS Abeta (+/-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.

PMID:
17383190
PMCID:
PMC2040126
DOI:
10.1016/j.jasms.2007.02.009
[Indexed for MEDLINE]
Free PMC Article

Publication types, MeSH terms, Substances, Grant support

Publication types

MeSH terms

Substances

Grant support

Supplemental Content

Full text links

Icon for Springer Icon for PubMed Central
Loading ...
Support Center