Asymmetry of the Active Site Loop Conformation between Subunits of Glutamate-1-semialdehyde Aminomutase in Solution.

Glutamate-1-semialdehyde aminomutase (GSAM) is a dimeric, pyridoxal 5'-phosphate (PLP)-dependent enzyme catalysing in plants and some bacteria the isomerization of L-glutamate-1-semialdehyde to 5-aminolevulinate, a common precursor of chlorophyll, haem, coenzyme B12, and other tetrapyrrolic compound...

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Publicado en:BioMed Research International Vol. 2013; pp. 353270 - 353271
Autores principales: Campanini, Barbara, Bettati, Stefano, di Salvo, Martino Luigi, Mozzarelli, Andrea, Contestabile, Roberto
Formato: Journal Article
Publicado: Wiley-Blackwell 2013
Acceso en línea:Ver este registro en EBSCOhost
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      jtl: BioMed Research International
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      dt: 2013
      vid: 2013
      pid: 480
      pub: Wiley-Blackwell
      place: Malden, Massachusetts
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        atl: Asymmetry of the Active Site Loop Conformation between Subunits of Glutamate-1-semialdehyde Aminomutase in Solution.
      aug:
        au:
          Campanini, Barbara
          Bettati, Stefano
          di Salvo, Martino Luigi
          Mozzarelli, Andrea
          Contestabile, Roberto
        affil: Dipartimento di Farmacia, Università di Parma, Parco Area delle Scienze 23/A, 43124 Parma, Italy.
      sug:
        subj:
          Amino Acids
          Enzymes
          Enzymes Metabolism
          Proteins
          Proteins Metabolism
          Gram-Negative Bacteria
          Absorption
          Biochemical Phenomena Drug Effects
          Coenzymes Metabolism
          Chemistry
          Potassium Iodide Pharmacodynamics
          Vitamin B Complex Metabolism
          Pyridoxine Metabolism
          Solutions
          Spectrometry, Fluorescence
      ab: Glutamate-1-semialdehyde aminomutase (GSAM) is a dimeric, pyridoxal 5'-phosphate (PLP)-dependent enzyme catalysing in plants and some bacteria the isomerization of L-glutamate-1-semialdehyde to 5-aminolevulinate, a common precursor of chlorophyll, haem, coenzyme B12, and other tetrapyrrolic compounds. During the catalytic cycle, the coenzyme undergoes conversion from pyridoxamine 5'-phosphate (PMP) to PLP. The entrance of the catalytic site is protected by a loop that is believed to switch from an open to a closed conformation during catalysis. Crystallographic studies indicated that the structure of the mobile loop is related to the form of the cofactor bound to the active site, allowing for asymmetry within the dimer. Since no information on structural and functional asymmetry of the enzyme in solution is available in the literature, we investigated the active site accessibility by determining the cofactor fluorescence quenching of PMP- and PLP-GSAM forms. PLP-GSAM is partially quenched by potassium iodide, suggesting that at least one catalytic site is accessible to the anionic quencher and therefore confirming the asymmetry observed in the crystal structure. Iodide induces release of the cofactor from PMP-GSAM, apparently from only one catalytic site, therefore suggesting an asymmetry also in this form of the enzyme in solution, in contrast with the crystallographic data.
      pubtype: Academic Journal
      doctype: Journal Article
      ougenre: Article
    language: English
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