Proteinase K: An Enzyme for Everyone

protein expression purification and analysisWe recently posted a blog about Proteinase K, a serine protease that exhibits broad cleavage activity produced by the fungus Tritirachium album Limber. It cleaves peptide bonds adjacent to the carboxylic group of aliphatic and aromatic amino acids and is useful for general digestion of protein in biological samples. In that previous blog we focused on its use to remove RNase and DNase activities. However, the stability of Proteinase K in urea and SDS and its ability to digest native proteins make it useful for a variety of applications. Here we provide a brief list of peer-reviewed citations that demonstrate the use of proteinase K in DNA and RNA purification, protein digestion in FFPE tissue samples, chromatin precipitation assays, and proteinase K protection assays:

Removal of proteins for DNA or RNA purification.

  1. Biran, J et al. (2012) Neurokinin Bs and neurokinin B receptors in zebrafish-potential role in controlling fish reproduction. Proc. Natl. Acad. 109, 10269–274.
  2. Guo, C. et al. (2012) Global identification of MLL2 –targeted loci reveal MML2 role in diverse signaling pathways. Proc. Natl. Acad. Sci. 109, 17603–608
  3. Martinez, F. et al. (2012) Leucine Zipper Domain is required for Kaposi Sarcoma-associated Herpesvirus K-bZIP Protein to interact with Histone Deacetylase and is important for KSHV Replication. J. Biol. Chem. 287, 15622–634.
  4. Lin, X. et al. (2012) MicroRNAs and Unusual Small RNAs Discovered in Kaposi’s Sarcoma-Associated Herpesvirus Virions. J. Vir. 86, 12717–730.

Protein digestion in formalin-fixed paraffin-embedded (FFPE) tissue.

  1. Bric, L. et al. (2012) Morphologic and Clinicopathologic Features of Lunf Squamous Cell Carcinomas Expressing Sox2. Amer. J. Clin. Path. 138, 712–18
  2. Wang, B. et al. (2012) RelB/NF-kappaB2 Regulates Corticotropin-Releasing Hormone in the Human Placenta. Mol. Endocrinol. 26,1356–69
  3. Zeng, M. et al. (2012) Critical role of CD4 T cells in maintaining lymphoid tissue structure for immune cell homeostatsis and reconstitution. Blood 120, 1856–67

Chromatin Immunoprecipitation Assay (CHIP)

  1. Zhao, L-F. et al. (2012) Liver X Receptor alpha is involved in the Transcriptional regulation of the 6-Phosphofructo-2-Kinase/Fructose -2,6-Bisphosphatase Gene. Diabetes 61, 1062–71
  2. He, X. et al. (2012) Redox Regulation of Nuclear Factor Erythroid 2 –Related Factor 2: Gatekeeping for the Basal and Diabetes-Induced Expression of Thioredoxin-Interacting Protein. Mol. Pharmacol. 82, 887–97
  3. Wang, B. et al. (2012) RelB/NF-kappaB2 Regulates Corticotropin-Releasing Hormone in the Human Placenta. Mol. Endocrinol. 26, 1356-69

Protease K Protection Assays (determination of protein translocation by the lipid bilayer of microsomal membranes)

  1. Minn, I. et al. (2009) SUN-1 and ZYG-12, mediators of centrosome-nucleus attachment, are a functional SUN/KASH pair in Caenorhabditis elegans. Mol. Biol. Cell. 20, 4586–95.
  2. Padhan, K. et al. (2007) Severe acute respiratory syndrome coronavirus Orf3a protein interacts with caveolin. J. Gen. Virol. 88, 3067–77.
  3. Tews, B.A. et al. (2007) The pestivirus glycoprotein Erns is anchored in plane in the membrane via an amphipathic helix. J. Biol. Chem. 282, 32730–41.
  4. Pidasheva, S. et al. (2005) Impaired cotranslational processing of the calcium-sensing receptor due to signal peptide missense mutations in familial hypocalciuric hypercalcemia. Hum. Mol. Gen. 14, 1679–90.

For additional information about Proteinase K Protection Assays, please see this blog post as well.

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Gary Kobs

Strategic Marketing Manager at Promega Corporation
Gary earned his B.S. in Bacteriology, UW-Madison in 1982. From 1982–1986 he served as Research Tech at UW-Madison. From 1986 to the present Gary has been with Promega Corporation serving in many capacities including as the very first editor of Promega Notes. He was also Manager Tech Services and Training, Product Manager Restriction/Modifying Enzymes, Product Manager Protein Analysis, and is now Marketing Manager Protein Analysis.

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