Recombinant Mycoplasma pneumoniae Uncharacterized lipoprotein MG186 homolog (MPN_133)
Description
Biochemical Properties of Recombinant MPN_133
Recombinant MPN_133 (rMpn133) retains enzymatic activity when expressed in Escherichia coli after codon optimization (e.g., UGA-to-TGG changes to preserve tryptophan residues) . Key biochemical traits include:
Deletion of the EKS region (rMpn133 Δ72–110) abolishes cellular internalization but does not impair nuclease activity .
Role in Pathogenesis
MPN_133 contributes to M. pneumoniae virulence through two mechanisms:
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Cytotoxicity: Induces apoptosis-like death in human airway (A549) cells, characterized by phosphatidylserine externalization and TUNEL-positive nuclei .
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Nutrient acquisition: Part of the mpn133-136 operon, potentially involved in glycerol or glycerophosphocholine (GPC) transport, though substrate specificity remains unresolved .
Recombinant Expression and Engineering
rMpn133 is cloned using primers designed for codon optimization and EKS region deletion (Table 1) .
Table 1: Primer Sequences for rMpn133 Cloning
| Primer Name | Sequence (5’→3’) | Purpose |
|---|---|---|
| MPN133 SF | GGAATTC CATATGACCCGCGACTATACTACCAAG | Amplification of mpn133 |
| MPNm66R | ATAGATGCGGAGACCGTGTCGGCGCTCTTCGCTTGC | EKS region deletion |
| MPN133 R1 | CGC GGATCCTTAACTACCCTTTTGGGCTAATTTG | Vector insertion |
Therapeutic Implications
MPN_133 is a target for bacterial chassis engineering. Deletion of mpn133 in attenuated M. pneumoniae strains (e.g., CV2 strain lacking mpn133 and mpn372) reduces cytotoxicity while maintaining viability in lung tissue . This modification enables engineered strains to serve as delivery systems for antimicrobial therapies .
Research Applications
Product Specs
Target Background
KEGG: mpn:MPN133
