RESUMEN

To identify sialic acid binding proteins from complex proteomes, three photocrosslinking affinity-based probes were constructed using Neu5Ac (5 and 6) and Neu5Ac2en (7) scaffolds. Kinetic inhibition assays and Western blotting revealed the Neu5Ac2en-based 7 to be an effective probe for the labeling of a purified gut microbial sialidase (BDI_2946) and a purified human sialic acid binding protein (hCD33). Additionally, LC-MS/MS affinity-based protein profiling verified the ability of 7 to enrich a low-abundance sialic acid binding protein (complement factor H) from human serum thus validating the utility of this probe in a complex context.

Asunto(s)

Inhibidores Enzimáticos/farmacología , Sondas Moleculares/farmacología , Ácido N-Acetilneuramínico/farmacología , Lectina 3 Similar a Ig de Unión al Ácido Siálico/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Humanos , Sondas Moleculares/síntesis química , Sondas Moleculares/química , Estructura Molecular , Ácido N-Acetilneuramínico/síntesis química , Ácido N-Acetilneuramínico/química , Neuraminidasa/antagonistas & inhibidores , Neuraminidasa/metabolismo , Lectina 3 Similar a Ig de Unión al Ácido Siálico/aislamiento & purificación , Lectina 3 Similar a Ig de Unión al Ácido Siálico/metabolismo , Relación Estructura-Actividad