TY - JOUR
T1 - Impedimetric Characterization of NanA Structural Domains Activity on Sialoside-Containing Interfaces
AU - Alshanski, Israel
AU - Toraskar, Suraj
AU - Mor, Karin
AU - Daligault, Franck
AU - Jain, Prashant
AU - Grandjean, Cyrille
AU - Kikkeri, Raghavendra
AU - Hurevich, Mattan
AU - Yitzchaik, Shlomo
N1 - Publisher Copyright:
© 2024 American Chemical Society.
PY - 2024
Y1 - 2024
N2 - Streptococcus pneumoniae is a pathogenic bacterium that contains the surface-bound neuraminidase, NanA. NanA has two domains that interact with sialosides. It is hard to determine the contribution of each domain separately on catalysis or binding. In this work, we used biochemical methods to obtain the separated domains, applied electrochemical and surface analysis approaches, and determined the catalytic and binding preferences toward a surface-bound library of sialosides. Impedimetric studies on two different surfaces revealed that protein-surface interactions provide a tool for distinguishing the unique contribution of each domain at the interface affecting the substrate preference of the enzyme in different surroundings. We showed that each domain has a sialoside-specific affinity. Furthermore, while the interaction of the sialoside-covered surface with the carbohydrate-binding domain results in an increase in impedance and binding, the catalytic domain adheres to the surface at high concentrations but retains its catalytic activity at low concentrations.
AB - Streptococcus pneumoniae is a pathogenic bacterium that contains the surface-bound neuraminidase, NanA. NanA has two domains that interact with sialosides. It is hard to determine the contribution of each domain separately on catalysis or binding. In this work, we used biochemical methods to obtain the separated domains, applied electrochemical and surface analysis approaches, and determined the catalytic and binding preferences toward a surface-bound library of sialosides. Impedimetric studies on two different surfaces revealed that protein-surface interactions provide a tool for distinguishing the unique contribution of each domain at the interface affecting the substrate preference of the enzyme in different surroundings. We showed that each domain has a sialoside-specific affinity. Furthermore, while the interaction of the sialoside-covered surface with the carbohydrate-binding domain results in an increase in impedance and binding, the catalytic domain adheres to the surface at high concentrations but retains its catalytic activity at low concentrations.
UR - http://www.scopus.com/inward/record.url?scp=85206446648&partnerID=8YFLogxK
U2 - 10.1021/acs.langmuir.4c02620
DO - 10.1021/acs.langmuir.4c02620
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C2 - 39376038
AN - SCOPUS:85206446648
SN - 0743-7463
JO - Langmuir
JF - Langmuir
ER -