This review will describe recent methodological strategies for studying protein O-mannosylation using in vitro and in vivo approaches. Protein O-mannosylation is evolutionarily conserved in metazoans, yet this pathway is simplified and more amenable to genetic analyses in invertebrate organisms, indicating that genetically tractable in vivo models could facilitate research in this area. However, this mechanism is underlain by complex genetic and molecular regulation that remain poorly understood. Recent progress in mass spectrometry and in vitro analyses has shed new light the mechanism of α-Dystroglycan glycosylation. Genetic defects of O-mannosylation result in the loss of ligand binding activity of α-Dystroglycan and causes congenital muscular dystrophies termed dystroglycanopathies. A major biological effect of O-mannosylation concentrates on the regulation of α-Dystroglycan, a membrane glycoprotein mediating cell – extracellular matrix interactions.
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