1H, 13C and 15N Assignment of Self-Complemented MrkA Protein Antigen from Klebsiella Pneumoniae
Biomolecular NMR Assignments(2024)
摘要
Abstract Klebsiella pneumoniae (Kp) poses an escalating threat to public health, particularly given its association with nosocomial infections and its emergence as a leading cause of neonatal sepsis, particularly in low- and middle-income countries (LMICs). Host cell adherence and biofilm formation of Kp is mediated by type 1 and type 3 fimbriae whose major fimbrial subunits are encoded by the fimA and mrkA genes, respectively. In this study, we focus on MrkA subunit, which is a 20 KDa protein whose 3D molecular structure remains elusive. We applied solution NMR to characterize a recombinant version of MrkA in which the donor strand segment situated at the protein's N-terminus is relocated to the C-terminus, preceded by a hexaglycine linker. This construct yields a self-complemented variant of MrkA. Remarkably, the self-complemented MrkA monomer loses its capacity to interact with other monomers and to extend into fimbriae structures. We succeed to assign the 13C-15N- self-complemented MrkA monomer, with carbon atoms resonances accounting for 81%, backbone nitrogen atoms resonances for 92%, and proton resonances for 92%. Furthermore, an examination of its internal mobility unveiled that relaxation parameters are predominantly uniform across the polypeptide sequence, except for the glycine-rich region within loop 175-180. These data pave the way to a comprehensive structural elucidation of the MrkA monomer and to structurally map the molecular interaction regions between MrkA and antigen-induced antibodies.
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关键词
NMR resonance assignment,mrkA,Klebsiella pneumoniae,Self-complemented monomer,Type 3 fimbriae,Heteronucelar relaxation data,Protein antigen
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