The homozygous M712T mutation of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase results in reduced enzyme activities but not in altered overall cellular sialylation in hereditary inclusion body myopathy |
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Authors: | Hinderlich Stephan Salama Ilan Eisenberg Iris Potikha Tamara Mantey Lars R Yarema Kevin J Horstkorte Rüdiger Argov Zohar Sadeh Menachem Reutter Werner Mitrani-Rosenbaum Stella |
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Affiliation: | Institute of Enzymology, Hungarian Academy of Sciences, Karolina u. 29, H-1113 Budapest, Hungary. |
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Abstract: | In vivo growth of bacterial flagellar filaments by self-assembly of flagellin is promoted by a capping structure composed of a pentameric assembly of hook associated protein 2 (HAP2). Isolated native filaments with intact HAP2 cap exhibited higher melting temperature (deltaTm = 4 degrees C) and significantly increased resistance against heat-induced depolymerization than non-capped ones. Reconstituted filaments were also stabilized by HAP2 binding, but the obtained filament-HAP2 complexes were less stable than native assemblies. Their fast depolymerization at elevated temperatures and sensitivity to proteolysis indicated that native-like filament-HAP2 complexes are rarely obtained by in vitro reconstitution. A procedure was developed to isolate perfectly capped native filaments to facilitate high-resolution structural analysis. |
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