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Von der Proteinexpressi on zur 3D-Röntgenstruktur

Von der Proteinexpressi on zur 3D-Röntgenstruktur. Ivo Tews Biochemiezentrum der Universität Heidelberg. pyridoxal kinase. pyridoxine (pyridoxol). pyridoxamine. pyridoxal. vitamin B 6 biosynthesis. transaminases. PMP pyridoxamine 5*-phosphate. PLP pyridoxal 5*-phosphate. Vitamin B 6.

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Von der Proteinexpressi on zur 3D-Röntgenstruktur

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  1. Von der Proteinexpression zur 3D-Röntgenstruktur Ivo Tews Biochemiezentrum der Universität Heidelberg

  2. pyridoxal kinase pyridoxine(pyridoxol) pyridoxamine pyridoxal vitamin B6 biosynthesis transaminases PMP pyridoxamine 5*-phosphate PLP pyridoxal 5*-phosphate Vitamin B6

  3. ammonia and glutamate are produced from glutamine glutamine vitamin B6 biosynthesis the enzyme catalysing this reaction is the glutaminase Pdx2 PLP is then synthesised from a triose, a pentose and ammonia Vitamin B6 biosynthesis the enzyme catalysing this reaction is the synthase Pdx1 • PLP synthesis occurs in two consecutive steps • these are linked • ammonia needs to be transferred

  4. glutamine ribose 5*-phosphate glyceraldehyde 3*-phosphate glutamine amidotransferases PLP synthase Pdx1 glutaminase Pdx2 linking glutaminase and synthase • PLP synthesis occurs in two consecutive steps • these are linked • ammonia needs to be transferred glutamine amidotransferase (GATase)

  5. glutamine amidotransferases linking glutaminase and synthase GATase I GATase II „triad“ class Ntn class Cys-His-Glu N-terminal nucleophile

  6. glutamine amidotransferases linking glutaminase and synthase GATase I GATase II „triad“ class Ntn class Cys-His-Glu N-terminal nucleophile

  7. glutamine amidotransferases linking glutaminase and synthase GATase I GATase II „triad“ class Ntn class Cys-His-Glu N-terminal nucleophile

  8. glutamine amidotransferases linking glutaminase and synthase GATase I GATase II „triad“ class Ntn class Cys-His-Glu N-terminal nucleophile

  9. amine product catalysed by the glutaminase ammonia glutamine enzyme tunnels catalysed by the synthase or ammonia acceptor domain

  10. ImGP synthase imidazoleglycerol phosphatesynthase bienzyme complex figure taken from Structure 10 (2002), 185 enzyme tunnels HisF HisH

  11. P. falciparum vs. B. subtilis Plasmodium falciparum Pdx2

  12. P. falciparum vs. B. subtilis Bacillus subtilis Pdx2

  13. Bacillus subtilis Pdx2 P. falciparum vs. B. subtilis Plasmodium falciparum Pdx2

  14. glutaminase activation Gly47 Gly47 Bacillus subtilis Pdx2 – two conformations in the crystal Cys79 glutamine substrate (model)

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