University of California

Sina Ghaemmaghami

John Newman

Jonathan S Weissman

Nicholas Ingolia

Stanley B Prusiner

Shenheng Guan

John C Price

Al Burlingame

Methods are provided for, inter alia, detecting nucleic acid molecules resistant to degradation, such as a plurality of RNA molecules bound to a ribosome, using various technologies including deep sequencing.

Methods for detecting modification resistant nucleic acids

An entire complement or plurality of isotopically labeled amino acids are introduced into the diet of a test subject. Sufficient amounts of the isotopically labeled amino acids are provided to the subject in order to ensure that the subject incorporates a large percentage of isotopically labeled amino acids into newly synthesized proteins. Tissue samples are removed from the subject at different points in time and proteins are extracted and separated so that different proteins of different tissues can be individually analyzed and their amount and pattern of isotopic labeling can be determined. In a preferred embodiment, the methodology can be combined with proteolytic digestion to peptides and analysis by mass spectrometry in order to measure rates of protein turnover in vivo relating to thousands of different proteins.

Isotopic labeling for the measurement of global protein levels and turnover in vivo

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San Francisco, CA, United States of America

Sina Ghaemmaghami