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Expression and purification of a MORN fusion protein from Pseudomonas aeruginosa
Author(s) -
Anderson Caity Cory,
MacDonald Logan C,
Nice Justin B.,
Berger Bryan W.,
Streu Craig N
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1021.11
Subject(s) - proteases , pseudomonas aeruginosa , protease , biology , microbiology and biotechnology , bacteria , immune system , immunology , enzyme , biochemistry , genetics
Pseudomonas aeruginosa is a Gram‐negative bacteria and opportunistic pathogen commonly associated with diseases such as corneal keratitis. While antibiotics can be effective at treating primary infections, secondary, host‐induced effects from chronic infection can still lead to major permanent scarring and possibly even blindness. Host‐induced tissue damage is a major complication associated with chronic P. aeruginosa infections, and paradoxically can in some instances enhance the severity of infection and bacterial persistence. This damage has been shown to be a result of the over‐activation of the host's proteases, including cathepsins and MMPs. Genomic screening of P. aeruginosa identified a putative bacterial protease, which may be involved in activating host immune proteases. This protease contains N‐terminal domains of substantial homology to membrane occupancy and recognition nexus (MORN), for which little structural information exists. We have demonstrated in transfected cells that the N‐terminus is required for optimal catalytic activity, which may suggest a role in host‐induced tissue damage. Herein we outline the successful bacterial expression and purification of this MORN domain using an MBP‐T7 fusion vector as well as outline an experimental approach for defining the role of this domain in activity of the putative protease and host induced tissue damage.