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bioRxiv · 10.1101/2022.06.07.495235

Serotype specific sugars impact structure but not functions of the trimeric autotransporter adhesin EmaA of Aggregatibacter actinomycetemcomitans.

TANG-SIEGEL, G.·Radermacher, M.·Mintz, K.·Ruiz, T.
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Abstract

The human oral pathobiont Aggregatibacter actinomycetemcomitans expresses multiple virulence factors including the trimeric, extracellular matrix protein adhesin A (EmaA). The posttranslational modification of EmaA is proposed to be dependent on the sugars and enzymes associated with O-polysaccharide (O-PS) synthesis of the lipopolysaccharide (LPS). This modification is important for the structure and function of this adhesin. To determine if the composition of the sugars alters structure and/or function, the prototypic 202 kDa protein was expressed in a non-serotype b, emaA mutant strain. The transformed strain displayed EmaA adhesins similar in appearance to the prototypic adhesin as observed by 2D electron microscopy of whole-mount negatively stained bacterial preparations. Biochemical analysis indicated that the protein monomers were post-translationally modified. 3D electron tomographic reconstruction and structure analyses of the functional domain revealed three well-defined subdomains (SI, SII and SIII) with a linker region between SII and SIII. Structural changes were observed in all three subdomains and the linker region of the adhesins synthesized compared with the known structure. These changes however did not affect the ability of the strain to bind collagen or form biofilms. The data suggest that changes in the composition of the glycan moiety alter the 3D structure of the molecule without negatively affecting the function(s) associated with this adhesin. IMPORTANCEThe human oral pathogen A. actinomycetemcomitans is a causative agent of periodontal and several systemic diseases. EmaA is a trimeric autotransporter protein adhesin important for the colonization of this pathobiont in vivo. This adhesin is modified with sugars associated with the O-polysaccharide (O-PS) and the modification is mediated using the same enzymes involved in lipopolysaccharide (LPS) biosynthesis. The interaction with collagen is not mediated by the specific binding between the glycans and collagen but is attributed to changes in the final quaternary structure necessary to maintain an active adhesin. In this study, we have determined that the composition of the sugars utilized in the post-translational modification of this adhesin is exchangeable without compromising functional activities.

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TANG-SIEGEL, G., Radermacher, M., Mintz, K., Ruiz, T.. 2022-06-08. Serotype specific sugars impact structure but not functions of the trimeric autotransporter adhesin EmaA of Aggregatibacter actinomycetemcomitans.. https://doi.org/10.1101/2022.06.07.495235

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