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Protein Acetylation

Volume 981 of the series Methods in Molecular Biology pp 151-165

Date:

Using Functional Proteome Microarrays to Study Protein Lysine Acetylation

  • Jin-ying LuAffiliated withDepartment of Laboratory Medicine, National Taiwan University Hospital, Institute of Molecular Medicine, College of Medicine, National Taiwan University
  • , Yu-yi LinAffiliated withDepartment of Oncology, National Taiwan University Hospital, Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University
  • , Jef D. BoekeAffiliated withDepartment of Molecular Biology and Genetics, The High Throughput Biology Center, John Hopkins University School of Medicine
  • , Heng ZhuAffiliated withDepartments of Pharmacology, The High Throughput Biology Center, John Hopkins University School of MedicineDepartment Molecular Sciences, The High Throughput Biology Center, John Hopkins University School of Medicine

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Abstract

Emergence of proteome microarray provides a versatile platform to globally explore biological functions of broad significance. In the past decade, researchers have successfully fabricated functional proteome microarrays by printing individually purified proteins at a high-throughput, proteome-wide scale on one single slide. These arrays have been used to profile protein posttranslational modifications, including phosphorylation, ubiquitylation, acetylation, and nitrosylation. In this chapter, we summarize our work of using the yeast proteome microarrays to connect protein lysine acetylation substrates to their upstream modifying enzyme, the nucleosome acetyltransferase of H4 (NuA4), which is the only essential acetyltransferase in yeast. We further prove that the reversible acetylation on critical cell metabolism-related enzymes controls life span in yeast. Our studies represent a paradigm shift for the functional dissection of a crucial acetylation enzyme affecting aging and longevity pathways.

Key words

Protein chip Posttranslational modification Lysine acetylation Functional proteome microarray Aging Longevity