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Revision as of 07:54, 17 October 2021

Outer Membrane Protein A (OmpA)

Description

Outer membrane protein A (OmpA) is a 35 kDa transmembrane protein found in many enterobacteria. It is responsible for maintaining the stability of the bacterial membrane and cell signaling. [1]

Biology

In E. coli, the β-barrel conformation of OmpA is composed of extracellular loops that help play a role in the detection of outer membrane stress and binding of extracellular molecules. OmpA also serves as a porin channel that regulates water transport within the bacterial cell. [2]

Usage

In our project, the sequence of OmpA protein was used to construct the OmpA/OprF chimeric protein.

To construct our OmpA/OprF chimeric protein, we replaced Loop 1 (AA 38-54) of OmpA from E. coli with Loop 5 (AA 198-237) of OprF from P. aeruginosa, [4] which resulted in the final OmpA/OprF chimeric protein. The new extracellular peptides of the OmpA/OprF chimeric protein allow for binding of IFN-γ to the E. coli outer membrane.

In the IFN-γ sensing system, OmpA/OprF chimeric protein expression is driven by the ompA promoter. Binding of IFN-γ to the OmpA/OprF chimeric protein activates the pspA promoter, producing the enzyme required for the synthesis of taurine.

Characterization

The ompA/oprF sequence was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.

Expression of OmpA/OprF chimeric protein was confirmed by western blot using anti-OmpA antibody.

Sequence and Features