Crystal structure of human glyoxalase II and its complex with a glutathione thiolester substrate analogue

Cameron, Alexander D.; Ridderström, Marianne; Olin, B.; Mannervik, Bengt

doi:10.1016/s0969-2126(99)80174-9

Cited by 192 publications

(330 citation statements)

References 55 publications

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“…Based on its similarity to the metallo-␤-lactamases, we predicted that glyoxalase II binds two Zn(II) ions, utilizing five histidines, two aspartic acids, and a bridging water molecule (18). These predictions have been restated by Melino et al (22) and supported through the recent determination of the human glyoxalase II crystal structure (23). The crystallographic data indicates that the metal binding and active sites of glyoxalase II resemble those in the metallo-␤-lactamase L1 from Stenotrophomonas maltophilia (24).…”

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confidence: 73%

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Arabidopsis Glyoxalase II Contains a Zinc/Iron Binuclear Metal Center That Is Essential for Substrate Binding and Catalysis

Zang

Hollman

Crawford

et al. 2001

Journal of Biological Chemistry

127

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Glyoxalase II participates in the cellular detoxification of cytotoxic and mutagenic 2-oxoaldehydes. Because of its role in chemical detoxification, glyoxalase II has been studied as a potential anti-cancer and/or antiprotozoal target; however, very little is known about the active site and reaction mechanism of this important enzyme. To characterize the active site and kinetic mechanism of the enzyme, a detailed mutational study of Arabidopsis glyoxalase II was conducted. Data presented here demonstrate for the first time that the cytoplasmic form of Arabidopsis glyoxalase II contains an iron-zinc binuclear metal center that is essential for activity. Both metals participate in substrate binding, transition state stabilization, and the hydrolysis reaction. Subtle alterations in the geometry and/or electrostatics of the binuclear center have profound effects on the activity of the enzyme. Additional residues important in substrate binding have also been identified. An overall reaction mechanism for glyoxalase II is proposed based on the mutational and kinetic data from this study and crystallographic data on human glyoxalase II. Information presented here provides new insights into the active site and reaction mechanism of glyoxalase II that can be used for the rational design of glyoxalase II inhibitors.The glyoxalase system consists of two enzymes that convert cytotoxic 2-oxoaldehydes into hydroxy acids utilizing the cofactor glutathione. Under physiological conditions, glyoxalase I (lactoylglutathione lyase) catalyzes the formation of S-D-lactoylglutathione (SLG) 1 in the presence of methylglyoxal and glutathione (1). Methylglyoxal, a cytotoxic compound that can inactivate proteins, modify guanylate residues in DNA, and create interstrand cross-links (2-4), is formed as a by-product of several biochemical reactions including that of triose-phosphate isomerase (2). Glyoxalase II (hydroxyacylglutathione hydrolase) hydrolyzes SLG to form D-lactic acid and regenerate glutathione (1). SLG is also known to exhibit cytotoxic effects through the inhibition of DNA synthesis (5). Therefore, the glyoxalase system is thought to play a major role in chemical detoxification (5, 6).The glyoxalase system has been the subject of intense study because of its potential implications in anti-protozoal and antitumor drug design (6 -8). Increased cellular levels of methylglyoxal and glyoxalase activity are associated with tumor cells, the malaria parasite, and several complications associated with diabetes (6). Inhibitors of glyoxalase I and glyoxalase II have been designed as potential anti-tumor agents; however, all inhibitors tested to date exhibited problems that limited their therapeutic usefulness (6 -9). It should be noted that these inhibition studies were all conducted with little or no information on the active site structure or the kinetic mechanism of the enzyme. Therefore, it is clear that a more detailed understanding of the active site of glyoxalase II and its reaction mechanism is essential to the rational design an...

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mentioning

confidence: 73%

“…It could not be purified, and therefore was not analyzed. Lysine 74 is located on the outside of the protein in the middle of a ␤ strand (23). The drastic change in protein solubility likely results from dramatic structural changes.…”

Section: Overexpression and Purification Of Wild Type And Mutantmentioning

confidence: 99%

Arabidopsis Glyoxalase II Contains a Zinc/Iron Binuclear Metal Center That Is Essential for Substrate Binding and Catalysis

Zang

Hollman

Crawford

et al. 2001

Journal of Biological Chemistry

127

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“…3(B)] was observed. The structures of A. thaliana and human GLX2s as well as other orthologs are composed of two tightly interacting domains: the N-terminal domain, which includes the first 174 residues and has the typical ab/ba fourlayer sandwich metallo-b-lactamase fold 16,17 and the smaller C-terminal domain is situated at one edge of the main core and consists of five helices. This domain is typical of the glyoxalase Type II family and was previously considered to be essential for substrate binding.…”

Section: Crystal Structure Of Ycblmentioning

confidence: 99%

“…This domain is typical of the glyoxalase Type II family and was previously considered to be essential for substrate binding. 16,17 GLX2-5 from A. thaliana mitochondria was crystallized as a dimer held together by a number of electrostatic interactions and with the C-termini buried in the dimeric interface. 18 However, it did not prove possible to confirm dimerization in solution, and thus it was concluded that it must be a crystallization artifact.…”

Section: Crystal Structure Of Ycblmentioning

confidence: 99%

“…Both histidine ligands form contacts through the Ne2 atom, in full agreement with the His-binding mode observed among the metallo-b-lactamase superfamily members. 11,17,20 One common structural feature of enzymes that have a metallo-b-lactamase fold is an extensive Hbonding network around the metal-binding ligands in the form of a so-called ''second shell'' of interactions. As YcbL represents the first example of a GLX2 enzyme with a single metal center, it is informative to compare the geometries of the secondshell amino acid metal ligands.…”

Section: Crystal Structure Of Ycblmentioning

confidence: 99%

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Structural and functional characterization of Salmonella enterica serovar Typhimurium YcbL: An unusual Type II glyoxalase

et al. 2010

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YcbL has been annotated as either a metallo-b-lactamase or glyoxalase II (GLX2), both members of the zinc metallohydrolase superfamily, that contains many enzymes with a diverse range of activities. Here, we report crystallographic and biochemical data for Salmonella enterica serovar Typhimurium YcbL that establishes it as GLX2, which differs in certain structural and functional properties compared with previously known examples. These features include the insertion of an ahelix after residue 87 in YcbL and truncation of the C-terminal domain, which leads to the loss of some recognition determinants for the glutathione substrate. Despite these changes, YcbL has robust GLX2 activity. A further difference is that the YcbL structure contains only a single bound metal ion rather than the dual site normally observed for GLX2s. Activity assays in the presence of various metal ions indicate an increase in activity above basal levels in the presence of manganous and ferrous ions. Thus, YcbL represents a novel member of the GLX2 family.

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Metallo β‐Lactamases

Herzberg

Fitzgerald

2004

Handbook of Metalloproteins

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The metallo‐β‐lactamases (class B β‐lactamases) confer bacterial resistance to β‐lactam antibiotics and are therefore targets for the development of clinical agents against resistant strains of bacteria. They are found in a wide variety of bacteria and act on a broad spectrum of substrates including penicillins, cephalosporins, and carbapenems, as well as on some inhibitors of serine‐β‐lactamases. These ∼25 000 Da zinc‐dependent enzymes have been studied extensively in recent years, and crystal structures have been determined for the enzymes from five different bacteria representing two different sequence subclasses (B1 and B3). The active site is located at the interface between two α/β domains where two zinc atoms bind. This review surveys the current understanding of these enzymes, with emphasis on structural studies, spectroscopy, kinetics, and proposals for the catalytic mechanism.

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Crystal structure of human glyoxalase II and its complex with a glutathione thiolester substrate analogue

Cited by 192 publications

References 55 publications

Arabidopsis Glyoxalase II Contains a Zinc/Iron Binuclear Metal Center That Is Essential for Substrate Binding and Catalysis

Arabidopsis Glyoxalase II Contains a Zinc/Iron Binuclear Metal Center That Is Essential for Substrate Binding and Catalysis

Structural and functional characterization of Salmonella enterica serovar Typhimurium YcbL: An unusual Type II glyoxalase

Metallo β‐Lactamases

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