Homogenization Does Not Alter nor Remove Post‐Translationally Modified Phosphorylations

Abstract

Post‐translational modification (PTM) describes the covalent and generally enzymatic modification of proteins during or after translation. Proteins undergo PTMs to form the mature protein product and are important components in cell signaling. For example, protein phosphorylation is a reversible post‐translational modification in which a phosphate group is covalently added to an amino acid residue by a protein kinase. Phosphorylation alters the structural conformation of a protein, which activates, deactivates, or modifies its function and is a target for many studies. Biological tissues are routinely homogenized in order to extract proteins, despite the concerns that the forces produced through homogenization may alter or remove PTMs. Here, using murine heart and kidney samples, we show that tissue homogenization of tissue does not alter nor remove post‐translationally modified phosphates. Furthermore, we show how detergents and sonication alter post‐translationally modified phosphate when compared to homogenization.Support or Funding InformationOmni International, Inc935 Cobb Place BlvdKennesaw Ga 30144