Structure of catalase HPII fromEscherichia coli at 1.9 � resolution

Abstract

Catalase HPII from Escherichia coli, a homotetramer of subunits with 753 residues, is the largest known catalase. The structure of native HPII has been refined at 1.9 A ˚ resolution using X-ray synchrotron data collected from crystals flash-cooled with liquid nitrogen. The crystallographic agreement factors R and R free are respectively 16.6% and 21.0%. The asymmetric unit of the crystal contains a whole molecule that shows accurate 222-point group symmetry. The structure of the central part of the HPII subunit gives a root mean square deviation of 1.5 A ˚ for 477 equivalencies with beef liver catalase. Most of the additional 276 residues of HPII are located in either an extended N-terminal arm or in a C-terminal domain organized with a flavodoxin-like topology. A small number of mostly hydrophilic interactions stabilize the relative orientation between the C-terminal domain and the core of the enzyme. The heme component of HPII is a cis-hydroxychlorin-spirolactone in an orientation that is flipped 180°with respect to the orientation of the heme found in beef liver catalase. The proximal ligand of the heme is Tyr415 which is joined by a covalent bond between its C atom and the N atom of His392. Over 2,700 well-defined solvent molecules have been identified filling a complex network of cavities and channels formed inside the molecule. Two channels lead close to the distal side heme pocket of each subunit suggesting separate inlet and exhaust functions. The longest channel , that begins in an adjacent subunit, is over 50 A ˚ in length, and the second channel is about 30 A ˚ in length. A third channel reaching the heme proximal side may provide access for the substrate needed to catalyze the heme modification and His-Tyr bond formation. HPII does not bind NADPH and the equivalent region to the NADPH binding pocket of bovine catalase, partially occluded in HPII by residues 585-590, corresponds to the entrance to the second channel. The heme distal pocket contains two solvent molecules, and the one closer to the iron atom appears to exhibit high mobility or low occupancy compatible with weak coordination. Proteins 1999;34:155-166.

The Crystal Structure of DJ-1, a Protein Related to Male Fertility and Parkinson's Disease

“…4, a and b) (20). The DALI search also revealed that DJ-1 has similar topology to three proteins: the domain of catalase HPII from E. coli (17,21); the subunit of anthranilate synthase TrpG from Sulfolobus solfataricus (22,23); and the domain of GMP synthetase from E. coli (Table II) (24). All of the three proteins have flavodoxin-like Rossmannfolds and belong to the Class I glutamine amidotransferase-like superfamily (GAT superfamily) involving thiJ domains (17).…”

Crystal structure of an intracellular protease fromPyrococcus horikoshiiat 2-Å resolution

“…The two proteins with the highest Z scores are the noncatalytic domain of E. coli catalase HPII which has no known function (18), and the glutamine amidotransferase (GA) domain of GMP synthetase (19). Fig.…”

Femtosecond resolution of ligand-heme interactions in the high-affinity quinol oxidase bd : A di-heme active site?

“…Specific channels conducting small substrates or product molecules like oxygen, hydrogen peroxide, and perhaps water to and from the heme active site have been discerned in many enzymes (ref. 43 and references therein), including cytochrome c oxidase aa 3 (39). More specifically, ferrous heme b 595 may provide a transient binding site for the ligand on its way from heme d to the medium in analogy to Cu B in the heme-copper oxidases (44).…”