The antibody to the denatured monomer of MOMP was designated A-5, while the antibody to the folded monomer was designated A-8

The antibody to the denatured monomer of MOMP was designated A-5, while the antibody to the folded monomer was designated A-8. Expression of in rMOMP was produced in using the pQE-30 vector in the QIAexpress expression Cav1.3 system (Qiagen). usually function as general or specific porins regulating the permeability of the membrane to small molecules (8, 9, 18, 36). Hence, the pore-forming molecules play an important part in the communication between bacteria and the environment. Structurally, a typical porin subunit consists of 16 to 18 -strands forming an antiparallel -barrel, with short turns located at the bottom of the barrel facing the periplasmic space and long loops located at the top end of the barrel facing the external surface of bacterial membrane (8, 9, 36, 42, 48). Three porin subunits are put together into a stable homotrimer, which is definitely highly resistant to detergent and protease, a key home required for enteric organisms to survive in the intestinal tract (8, 9). Experimentally, porins can be recognized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in three different conformational forms including the folded monomer, the denatured monomer, and the native trimer. Notably, different bacterial porins are amazingly related in their -barrel structure, although their main sequences have little homology (8, 12, 18, 36). As the major components of the gram-negative bacterial outer membrane, pore-forming proteins also play a role in bacterial pathogenesis, such as adherence, invasion, Cefazolin Sodium and serum resistance (35, 44C46). Porin-based vaccines can induce protecting immunity Cefazolin Sodium against some bacterial infections (39, 49). is the leading bacterial cause of human enteritis in many industrialized countries (20, 41). This pathogenic organism causes watery diarrhea and/or hemorrhagic colitis and is also associated with Guillain-Barr syndrome, an acute flaccid paralysis that may lead to respiratory muscle mass compromise and death (31, 40). The majority of human infections are epidemiologically linked to ingestion Cefazolin Sodium of contaminated food or water (20, 41). Despite the recent advances in our understanding of the pathobiology of colonizes and adapts to numerous sponsor or environmental conditions are still poorly understood. As with other pathogenic organisms, membrane surface molecules of are considered the major mediators of the pathogen-host relationships. The OMP profile of thermophilic varieties (including and has a molecular mass of 40 to 48 kDa in different strains (5, 16). Manifestation of MOMP seems stable under a variety of growth, incubation, and passage conditions (5). Based on its biochemical and structural features, MOMP has been characterized as a member of the trimeric bacterial porin superfamily (7, 16). Purified MOMP from offers pore-forming activity when reconstituted in lipid bilayers (10, 51). Much like additional gram-negative bacterial porins, MOMP exhibits heat modifiability and may be recognized in three conformational forms including the folded monomer (35 kDa), the denatured monomer (40 to 48 kDa), and the native trimer (120 to 140 kDa) (16). The trimeric form of MOMP has been observed within the membrane surface of cells (1, 2) and is less stable than the porin trimers of (7). In addition to being a porin, MOMP may have functions unique from its pore-forming activity. Purified MOMP bound to isolated membranes of the INT 407 cell membrane and partially inhibited Cefazolin Sodium the attachment of to sponsor cells (30, 37). The binding was apparently dependent on the conformation of MOMP, since the activity was observed only with MOMP purified under native conditions, not with MOMP purified with SDS. In a recent study by Bacon et al.(3), a cytotoxic complex composed of MOMP and lipopolysaccharide was identified and characterized from a clinical isolate of to many hydrophilic antibiotics (33)..