Christopher Petzold and Alyssa Redding-Johanson were key members of a team that demonstrated a technique called “targeted proteomics” to speed up and improve the identification and quantification of proteins within a sample. Efforts to engineer new metabolic pathways into microbes for the inexpensive production of valuable chemical products, such as biofuels or therapeutic drugs, should get a significant boost in a new development from the U.S. Petzold, who also holds an appointment with the Lawrence Berkeley National Laboratory (Berkeley Lab)’s Physical Biosciences Division, is the corresponding author on a paper describing this research that was published in the journal Metabolic Engineering.
Metabolic engineering is the practice of altering genes and chemical pathways within a cell or microorganism to increase production of a specific chemical substance.
Researchers have tried developing assays to evaluate every intermediate in a given metabolic pathway but this has been a major challenge because intermediates often degrade rapidly or are isomers, and because there are few available standards for such evaluations. Targeted proteomics is based on a variation of mass spectrometry called selected-reaction monitoring (SRM) that can be used to rapidly detect and quantify multiple target proteins within complex protein mixtures, such as those found in cells or microbes. Schematic of targeted proteomics technique in which a peptide mass and a specific fragment mass are selected for SRM mass spectrometry analysis to identify and quantify a target protein. Petzold and his co-authors demonstrated the effectiveness of their targeted proteomics technique when they used it to measure protein levels in Escherichia coli that were engineered with yeast proteins to produce amorpha-4,11-diene, a member of the family of plant chemicals known as sesquiterpenes. Our analysis identified two mevalonate pathway proteins, mevalonate kinase and phosphomevalonate kinase, both from yeast, as potential bottlenecks,” Petzold says.
Petzold and his JBEI colleagues are now in the process of implementing “scheduled-SRM,” a variation of the SRM mass spectrometry analysis that would allow them to easily detect and quantify 100 proteins in a single experiment that can be completed in less than two hours. Lawrence Berkeley National Laboratory addresses the world’s most urgent scientific challenges by advancing sustainable energy, protecting human health, creating new materials, and revealing the origin and fate of the universe. If you are looking to get a six pack fast, there is a revolutionary six pack ab workout program that is taking the fitness world by storm. We will show you how to get a six pack fast, with our comprehensive hot six pack ab workout GUARANTEED!!
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This is made possible by selecting only two points of mass for monitoring – a peptide mass and a specific fragment mass – rather than scanning the entire mass range. It is a scientific partnership led by Berkeley Lab and including the Sandia National Laboratories, the University of California campuses of Berkeley and Davis, the Carnegie Institution for Science, and the Lawrence Livermore National Laboratory.
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