Structure of quinoprotein methylamine dehydrogenase at 2.25 A resolution

• Published in: The EMBO journal 1989-08, Vol.8 (8), p.2171-2178
• Main Authors: Vellieux, F.M., Huitema, F., Groendijk, H., Kalk, K.H., Jzn, J.F., Jongejan, J.A., Duine, J.A., Petratos, K., Drenth, J., Hol, W.G.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Binding Sites; Biological and medical sciences; Chemical Phenomena; Chemistry; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Molecular Sequence Data; Molecular Structure; Molecular Weight; Neuraminidase; Oxidoreductases; Oxidoreductases Acting on CH-NH Group Donors - analysis; Protein Conformation; Thiobacillus - enzymology; X-Ray Diffraction
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1002/j.1460-2075.1989.tb08339.x

The three‐dimensional structure of quinoprotein methylamine dehydrogenase from Thiobacillus versutus has been determined at 2.25 A resolution by a combination of multiple isomorphous replacement, phase extension by solvent flattening and partial structure phasing using molecular dynamics refinement. In the resulting map, the polypeptide chain for both subunits could be followed and an X‐ray sequence was established. The tetrameric enzyme, made up of two heavy (H) and two light (L) subunits, is a flat parallellepiped with overall dimensions of approximately 76 x 61 x 45 A. The H subunit, comprising 370 residues, is made up of two distinct segments: the first 31 residues form an extension which embraces one of the L subunits; the remaining residues are found in a disc‐shaped domain. This domain is formed by a circular arrangement of seven topologically identical four‐stranded antiparallel beta‐sheets, with approximately 7‐fold symmetry. In spite of distinct differences, this arrangement is reminiscent of the structure found in influenza virus neuraminidase. The L subunit consists of 121 residues, out of which 53 form a beta‐sheet scaffold of a central three‐stranded antiparallel sheet flanked by two shorter two‐stranded antiparallel sheets. The remaining residues are found in segments of irregular structure. This subunit is stabilized by six disulphide bridges, plus two covalent bridges involving the quinone co‐factor and residues 57 and 107 of this subunit. The active site is located in a channel at the interface region between the H and L subunits, and the electron density in this part of the molecule suggests that the co‐factor of this enzyme is not pyrrolo quinoline quinone (PQQ) itself, but might be instead a precursor of PQQ.