Part:BBa_K4200000
Multivalent Adhesion Molecule 7
The part encodes the adhesion molecule MAM7 lacking the first 44 amino acids which forms a transmembrane helix. It is derived from Vibrio parahaemolyticus, where the native protein is present in its membrane and is used by the bacteria for mediating initial adhesion to host cell. The truncated coding region produces protein that remains cytosolic. The cytosolic protein can be purified and used to test the interaction of the adhesion molecule with any molecule that has been proposed to inhibit its interactions with its native ligand Fibronectin.
Usage and Biology
The native protein, from which the part is derived, contains an N-terminus transmembrane helix followed by seven tandem mammalian cell entry(MCE) domains. The MCE domains were determined to interact with the Phosphatidic acid and the Heparin Binding domain of Fibronectin. Furthermore, it has been established that while any of the MCE domains can bind the PA, at least five tandem repeats of MCE domains are required for interaction with Fibronectin. The interaction between the MAM7 molecule and the host cell ligands was found to be crucial for initiating host cell adhesion and, subsequently, the infection cycle. Our part consists of the portion of MAM7 encoding the MCE domains. It was reported that the truncated protein can still mediate the reported interactions, and thus the derived protein has been used as an inhibitor of the Gram-negative pathogen Staphylococcus aureus. However, another study reported that the MAM7 molecule, when it binds to Phosphatidic acid, can induce actin rearrangement, eventually disrupting epithelial integrity. This rules out the use of MAM7 as a potential inhibitor of infection.
We designed the part so it could be used to produce the pure protein, which could then be used to assay the interaction of the proposed antimicrobial peptide.
References
A. M. Krachler, H. Ham, and K. Orth, “Outer membrane adhesion factor multivalent adhesion molecule 7 initiates host cell binding during infection by gram-negative pathogens,” Proceedings of the National Academy of Sciences, vol. 108, no. 28, pp. 11614–11619, 2011.
C. Hawley, C. Watson, K. Orth and A. Krachler, "A MAM7 Peptide-Based Inhibitor of Staphylococcus aureus Adhesion Does Not Interfere with In Vitro Host Cell Function", PLoS ONE, vol. 8, no. 11, p. e81216, 2013.
J. Lim, D. Stones, C. Hawley, C. Watson and A. Krachler, "Multivalent Adhesion Molecule 7 Clusters Act as Signaling Platform for Host Cellular GTPase Activation and Facilitate Epithelial Barrier Dysfunction", PLoS Pathogens, vol. 10, no. 9, p. e1004421, 2014.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 126
Illegal BamHI site found at 1141
Illegal XhoI site found at 2521 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 46
Illegal NgoMIV site found at 867
Illegal NgoMIV site found at 1317
Illegal AgeI site found at 1264 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 304
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