Deoxyhypusine Hydroxylase Is an Fe(II)-dependent, Heat-repeat Enzyme

Kim, So Hyun; Kang, Kee Ryeon; Wolff, Edith C.; Bell, Jessica K.; McPhie, Peter; Park, Myung Hee

doi:10.1074/jbc.m601081200

Cited by 50 publications

(43 citation statements)

References 44 publications

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“…Enzymes-Human recombinant mutant DOHH enzymes with a single amino acid of the strictly conserved His-Glu motifs replaced with alanine (H56A, E57A, H89A, E90A, H207A, E208A, H240A, and E241A) were reported previously (24). 28 additional mutant enzymes with alanine substitution of all other conserved amino acids (R26A, L28A, K55A, G63A, Q64A, L74A, R88A, E93A, E120A, T121A, C122A, D148A, P149A, R175A, Y176A, R183A, S202A, V212A, G214A, Q215A, L225A, E234A, M237A, R239A, I246A, G247A, I258A, and S272A) and those with substitutions of Glu 57 or Glu 208 with aspartic acid, asparagine, or glutamine, respectively (E57D, E57N, E57Q, E208D, E208N, and E208Q), were generated using the QuikChange Site-directed Mutagenesis Kit.…”

Section: Generation Of Dohh Mutant Enzymes and Truncatedmentioning

confidence: 99%

“…In many of the double-stranded ␤ helix enzymes, iron is coordinated by His-X-Asp/Glu-X n -His at three coordination sites, and by a 2-oxoacid and oxygen (22). Judging from the stoichiometry of 2 mol of iron per mol of DOHH holoenzyme, DOHH probably contains one binuclear (diiron) active center (24 (24). As for the two mutant enzymes E57A and E208A, which were totally inactive despite their normal iron content, the reason for their lack of activity was unknown.…”

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confidence: 99%

“…The iron-binding active site of DOHH and mode of iron binding also differ from those of 2-oxoacid and Fe(II)-dependent dioxygenases, suggesting a distinct reaction mechanism (24). In many of the double-stranded ␤ helix enzymes, iron is coordinated by His-X-Asp/Glu-X n -His at three coordination sites, and by a 2-oxoacid and oxygen (22).…”

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confidence: 99%

“…DOHH consists of eight HEAT repeats, in a symmetrical dyad of four HEAT motifs connected by a variable region. The predicted ␣-helical structure of DOHH was validated experimentally by determination of the ␣-helical content of purified human recombinant DOHH by CD analysis (77 versus 76 -78%, calculated from the HEAT repeat model) (24).…”

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confidence: 99%

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Specificity of the Deoxyhypusine Hydroxylase-Eukaryotic Translation Initiation Factor (eIF5A) Interaction

Kang¹,

Kim²,

Wolff

et al. 2007

Journal of Biological Chemistry

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Deoxyhypusine hydroxylase (DOHH) is a novel metalloenzyme that catalyzes the final step of the post-translational synthesis of hypusine (N ⑀ -(4-amino-2-hydroxybutyl)lysine) in the eukaryotic translation initiation factor 5A (eIF5A). Hypusine synthesis is unique in that it occurs in only one protein, denoting the strict specificity of the modification enzymes toward the substrate protein. The specificity of the interaction between eIF5A and DOHH was investigated using human eIF5A (eIF5A-1 isoform) and human recombinant DOHH. DOHH displayed a strong preference for binding the deoxyhypusine-containing form of eIF5A, over the eIF5A precursor or the hypusine-containing eIF5A, indicating a role for the deoxyhypusine residue in binding. In addition to the deoxyhypusine residue, a large portion of the eIF5A polypeptide (>20 -90 amino acids) is required for effective modification by DOHH. We have identified the amino acid residues of DOHH that are critical for substrate binding by alanine substitution of 36 conserved amino acid residues. Of these, alanine substitution at Glu 57 is an unusual amino acid that occurs in all eukaryotes. It is formed posttranslationally in eukaryotic translation initiation factor 5A (eIF5A) 4 through the action of two enzymes (for a recent review, see Ref. 1). The first enzyme, deoxyhypusine synthase (DHS), catalyzes the NAD-dependent cleavage of spermidine and transfer of its 4-aminobutyl moiety to the ⑀-amino group of a specific lysine residue to form the deoxyhypusine (N ⑀ -(4-aminobutyl)lysine) residue (2, 3). The second enzyme, deoxyhypusine hydroxylase (DOHH), hydroxylates this intermediate to form the hypusine residue and mature eIF5A (4, 5).eIF5A and its modification enzymes, DHS and DOHH, are essential for mammalian cell proliferation (1, 6 -11). Various metal chelating inhibitors of DOHH, including mimosine and ciclopirox olamine, inhibit cell proliferation by causing cell cycle arrest at the G 1 /S boundary (12). Ciclolopirox olamine, in particular, exerted strong inhibition of human umbilical vein endothelial cell (HUVEC) proliferation and angiogenesis in model assays, suggesting the potential utility of DOHH inhibitors as antitumor agents (13). Although DOHH has been proposed as a potential target of antitumor therapy (13) and antihuman immunodeficiency virus type 1 therapy (14), no specific inhibitors of DOHH are currently available.The unique feature of hypusine formation is the strict substrate specificity of this protein modification. Hypusine synthesis occurs exclusively in one cellular protein, eIF5A precursor, at one specific lysine residue (Lys 50 for the human protein) and thereby represents one of the most specific protein modifications known to date. eIF5A and its modifying enzymes, DHS and DOHH, are highly conserved in all eukaryotes (1,8). The amino acid sequence conservation surrounding the hypusine site of eIF5A is especially high in eukaryotes, suggesting that the hypusine residue has an important basic function that has been preserved through evolution. Furthe...

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Section: Generation Of Dohh Mutant Enzymes and Truncatedmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

mentioning

confidence: 99%

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Specificity of the Deoxyhypusine Hydroxylase-Eukaryotic Translation Initiation Factor (eIF5A) Interaction

Kang¹,

Kim²,

Wolff

et al. 2007

Journal of Biological Chemistry

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“…A model of Cpc7 threaded onto known HEAT repeat proteins using Phyre2 indicates that Cpc7 is made of 27 tandem alpha helices (45). Proteins containing HEAT repeats can vary in function and have been described as having roles in scaffolding (36), lyase function (34,35), iron binding (37), and [Fe-S] cluster stabilization (38,39). While the mechanism of Cpc7-mediated regulation of development in M. xanthus is still unknown, we predict that it is involved in production of a developmental signal.…”

Section: Discussionmentioning

confidence: 99%

Chemosensory Regulation of a HEAT-Repeat Protein Couples Aggregation and Sporulation in Myxococcus xanthus

et al. 2014

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cChemosensory systems are complex, highly modified two-component systems (TCS) used by bacteria to control various biological functions ranging from motility to sporulation. Chemosensory systems and TCS both modulate phosphorelays comprised of histidine kinases and response regulators, some of which are single-domain response regulators (SD-RRs) such as CheY. In this study, we have identified and characterized the Che7 chemosensory system of Myxococcus xanthus, a common soil bacterium which displays multicellular development in response to stress. Both genetic and biochemical analyses indicate that the Che7 system regulates development via a direct interaction between the SD-RR CheY7 and a HEAT repeat domain-containing protein, Cpc7. Phosphorylation of the SD-RR affects the interaction with its target, and residues within the ␣4-␤5-␣5 fold of the REC domain govern this interaction. The identification of the Cpc7 interaction with CheY7 extends the diversity of known targets for SD-RRs in biological systems.

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Iron: Non‐Heme Proteins with Diiron‐Carboxylate Active Sites

Kurtz

Boice

Caranto

et al. 2004

Encyclopedia of Inorganic and Bioinorganic Chemistry

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Non‐heme diiron‐carboxylate ( NHDC ) active sites are found in several classes of monooxygenases, oxidases, and dioxygen transport or sensing proteins, as well as a few metallohydrolases. The unique and defining structural feature of NHDC sites consists of two non‐heme irons bridged by at least one carboxylate‐containing amino acid residue. A second characteristic feature is a bridging solvent ligand, either oxo or hydroxo in at least the diferric state. NHDC sites also feature terminal ligands derived from histidine and carboxylate residues. The hemerythrin superfamily functions have expanded from reversible dioxygen binding to include sensing of iron, dioxygen, or redox status. Substantial progress has been made in characterization of flavo‐diiron enzymes, including its nitric oxide reductase mechanism. A diiron urease is a recent addition to the metallohydrolase family. The variety of characterized NHDC O 2 ‐activating enzymes has increased substantially since the previous compilation in this encyclopedia. Novel monooxygenation mechanisms have been identified.

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Deoxyhypusine Hydroxylase Is an Fe(II)-dependent, Heat-repeat Enzyme

Cited by 50 publications

References 44 publications

Specificity of the Deoxyhypusine Hydroxylase-Eukaryotic Translation Initiation Factor (eIF5A) Interaction

Specificity of the Deoxyhypusine Hydroxylase-Eukaryotic Translation Initiation Factor (eIF5A) Interaction

Chemosensory Regulation of a HEAT-Repeat Protein Couples Aggregation and Sporulation in Myxococcus xanthus

Iron: Non‐Heme Proteins with Diiron‐Carboxylate Active Sites

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