Analysis of Bacterial Membrane Proteins Produced During Mammalian Infection Using Hydrophobic Antigen Tissue Triton Extraction (HATTREX)

Abstract

Hydrophobic antigen tissue Triton extraction (HATTREX) provides a method to extract and identify hydrophobic bacterial membrane proteins from host-infected tissues. The non-ionic detergent Triton X-114 is used to solubilize host-infected tissues at 4 degrees C. Subsequent phase partitioning of Triton X-114 extracted material at 37 degrees C results in a "detergent poor" aqueous phase and a "detergent rich" detergent phase. While soluble proteins partition to the aqueous phase, hydrophobic proteins, such as proteins of the outer and inner membranes, can be found in the detergent phase. Characterization of the detergent phase sample provides insights into the proteome of bacterial membranes during infection.