DOI: 10.1111/j.1432-1033.1995.372_1.x. Hydrogen Isotope Effects in the Reactions Catalyzed by H2‐forming N5,N10‐Methylenetetrahydromethanopterin Dehydrogenase from Methanogenic Archaea

Hydrogen Isotope Effects in the Reactions Catalyzed by H2‐forming N5,N10‐Methylenetetrahydromethanopterin Dehydrogenase from Methanogenic Archaea

10.1111/j.1432-1033.1995.372_1.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

H2‐forming N5,N10‐methylenetetrahydromethanopterin dehydrogenase from methanogenic Archaea, which is a novel hydrogenase containing neither nickel nor iron‐sulfur clusters, catalyzes the reversible reduction of N5,N10‐methenyltetrahydomethanopterin (CH≡H4MPT4) with H2 to N5,N10‐methylenetetrahydromethanopterin (CH2=H4MPT) and a proton (ΔG°′=− 5.5 kJ/mol). The enzyme also catalyzes a CH≡H4MPT+‐dependent H2/H+ exchange. We report here on kinetic deuterium isotope effects in these reactions. When CH≡H4MPT+ reduction was performed with D2 instead of H2, Vmax and the Km did not change. A primary isotope effect of 1 was found at all pH and temperatures tested and independent of whether H2O or D2O was the solvent. The findings indicate that a step other than the activation of H2 was rate‐determining in CH≡H4MPT+ reduction with H2. This was substantiated by the observation that also the CH≡H4MPT+‐dependent H2/H+ exchange reaction did not exhibit an appreciable deuterium isotope effect. VMax for CH2=H4MPT dehydrogenation to CH≡H4MPT+ and H2 was only 2–3 times higher than for CD2=H4MPT dehydrogenation to CD≡H4MPT+ and HD. Such a small primary isotope effect indicates that the breakage of the C‐H bond in the methylene group of CH2=H4MPT was only rate‐limiting when hydrogen was substituted by a deuterium.

Bibliography

Klein, A. R., Hartmann, G. C., & Thauer, R. K. (1995). Hydrogen Isotope Effects in the Reactions Catalyzed by H2âforming N5,N10âMethylenetetrahydromethanopterin Dehydrogenase from Methanogenic Archaea. European Journal of Biochemistry, 233(1), 372â376. Portico.

Authors 3

Andreas R. Klein (first)
Gudrun C. Hartmann (additional)
Rudolf K. Thauer (additional)

References 28 Referenced 30

10.1016/0005-2728(94)90036-1
10.1002/ange.19951072011 / Angew. Chem. / Zum Katalysemechanismus der Nickel‐ und Eisen‐Cluster‐freien Hydrogenase aus methanogenen Archaea: Enzymatische H2‐Aktivierung ohne Metall und ihre Analogie zur Chemie der Alkane in supersaurer Lösung by Berkessel A. (1995)
{'key': 'e_1_2_3_4_1', 'first-page': '634', 'article-title': 'Chemistry and spectroscopy in strongly acidic solutions: Reversible reaction between alipathic carbonium ions and hydrogen', 'volume': '67', 'author': 'Bickel A. F.', 'year': '1967', 'journal-title': 'J. Chem. Soc., Chem. Commun.'} / J. Chem. Soc., Chem. Commun. / Chemistry and spectroscopy in strongly acidic solutions: Reversible reaction between alipathic carbonium ions and hydrogen by Bickel A. F. (1967)
10.1111/j.1432-1033.1992.tb17502.x
{'key': 'e_1_2_3_6_1', 'first-page': '1', 'volume-title': 'The enzymes', 'author': 'Cleland W. W.', 'year': '1970'} / The enzymes by Cleland W. W. (1970)
{'key': 'e_1_2_3_7_1', 'volume-title': 'Enzyme mechanism from isotope effects', 'author': 'Cook P. F.', 'year': '1991'} / Enzyme mechanism from isotope effects by Cook P. F. (1991)
{'key': 'e_1_2_3_8_1', 'first-page': '203', 'volume-title': 'Enzyme mechanism from isotope effects', 'author': 'Cook P. F.', 'year': '1991'} / Enzyme mechanism from isotope effects by Cook P. F. (1991)
10.1021/ac60260a013
10.1016/S0021-9258(17)42721-9
Hartmann G. C.(1994)H2‐bildendeN5 N10‐Methylentetrahydromethanopterin‐Dehydrogenase und andere Enzyme der Methanogenese ausMethanococcus thermolithotrophicus Diploma Thesis Philipps‐Universität Marburg.
10.1128/jb.143.2.628-636.1980
10.1007/BF00418186
10.1007/BF00644273
10.1128/jb.177.9.2460-2468.1995
10.1002/ange.19931050604
{'key': 'e_1_2_3_17_1', 'first-page': '74', 'volume-title': 'Enzyme mechanism from isotope effects', 'author': 'Quinn D. M.', 'year': '1991'} / Enzyme mechanism from isotope effects by Quinn D. M. (1991)
{'key': 'e_1_2_3_18_1', 'first-page': '321', 'volume-title': 'The enzymes', 'author': 'Richards J. H.', 'year': '1970'} / The enzymes by Richards J. H. (1970)
10.1016/0014-5793(92)81522-N
10.1021/bi00179a027
10.1021/ja00115a035
10.1007/BF00244962
10.1111/j.1432-1033.1993.tb17657.x
10.1007/978-1-4615-2391-8_5
Vaupel M.(1993)Untersuchungen mitMethanobacterium thermoformicicumundMethanobacterium formicicumzur Rolle von H2‐bildenderN5 N10‐Methylentetrahydromethanopterin Dehydrogenase in methanogenen Archaea. Diploma Thesis Philipps‐Universität Marburg.
10.1111/j.1432-1033.1991.tb16491.x
10.1016/0092-8674(93)90570-G
10.1016/0014-5793(90)80649-4
10.1111/j.1432-1033.1992.tb17215.x

Dates

Type	When
Created	20 years, 11 months ago (Sept. 8, 2004, 4:04 a.m.)
Deposited	1 year, 9 months ago (Oct. 31, 2023, 8:56 a.m.)
Indexed	3 months, 1 week ago (May 18, 2025, 10:24 a.m.)
Issued	29 years, 10 months ago (Oct. 1, 1995)
Published	29 years, 10 months ago (Oct. 1, 1995)
Published Online	20 years, 11 months ago (Sept. 8, 2004)
Published Print	29 years, 10 months ago (Oct. 1, 1995)

Funders 0

None

BibTeX

@article{Klein_1995, title={Hydrogen Isotope Effects in the Reactions Catalyzed by H2‐forming N5,N10‐Methylenetetrahydromethanopterin Dehydrogenase from Methanogenic Archaea}, volume={233}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1995.372_1.x}, DOI={10.1111/j.1432-1033.1995.372_1.x}, number={1}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Klein, Andreas R. and Hartmann, Gudrun C. and Thauer, Rudolf K.}, year={1995}, month=oct, pages={372–376} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        5,
        18
      ]
    ],
    "date-time": "2025-05-18T14:24:43Z",
    "timestamp": 1747578283846
  },
  "reference-count": 28,
  "publisher": "Wiley",
  "issue": "1",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2004,
            9,
            8
          ]
        ],
        "date-time": "2004-09-08T00:00:00Z",
        "timestamp": 1094601600000
      },
      "content-version": "vor",
      "delay-in-days": 3265,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1995,
        10
      ]
    ]
  },
  "abstract": "<jats:p>H<jats:sub>2</jats:sub>\u2010forming <jats:italic>N</jats:italic><jats:sup>5</jats:sup>,<jats:italic>N</jats:italic><jats:sup>10</jats:sup>\u2010methylenetetrahydromethanopterin dehydrogenase from methanogenic Archaea, which is a novel hydrogenase containing neither nickel nor iron\u2010sulfur clusters, catalyzes the reversible reduction of <jats:italic>N</jats:italic><jats:sup>5</jats:sup>,<jats:italic>N</jats:italic><jats:sup>10</jats:sup>\u2010methenyltetrahydomethanopterin (CH\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>4</jats:sup>) with H<jats:sub>2</jats:sub> to N<jats:sup>5</jats:sup>,N<jats:sup>10</jats:sup>\u2010methylenetetrahydromethanopterin (CH<jats:sub>2</jats:sub>=H<jats:sub>4</jats:sub>MPT) and a proton (\u0394G\u00b0\u2032=\u2212 5.5 kJ/mol). The enzyme also catalyzes a CH\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>+</jats:sup>\u2010dependent H<jats:sub>2</jats:sub>/H<jats:sup>+</jats:sup> exchange. We report here on kinetic deuterium isotope effects in these reactions. When CH\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>+</jats:sup> reduction was performed with D<jats:sub>2</jats:sub> instead of H<jats:sub>2</jats:sub>, <jats:italic>V</jats:italic><jats:sub>max</jats:sub> and the <jats:italic>K</jats:italic><jats:sub>m</jats:sub> did not change. A primary isotope effect of 1 was found at all pH and temperatures tested and independent of whether H<jats:sub>2</jats:sub>O or D<jats:sub>2</jats:sub>O was the solvent. The findings indicate that a step other than the activation of H<jats:sub>2</jats:sub> was rate\u2010determining in CH\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>+</jats:sup> reduction with H<jats:sub>2</jats:sub>. This was substantiated by the observation that also the CH\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>+</jats:sup>\u2010dependent H<jats:sub>2</jats:sub>/H<jats:sup>+</jats:sup> exchange reaction did not exhibit an appreciable deuterium isotope effect. <jats:italic>V</jats:italic><jats:sub>Max</jats:sub> for CH<jats:sub>2</jats:sub>=H<jats:sub>4</jats:sub>MPT dehydrogenation to CH\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>+</jats:sup> and H<jats:sub>2</jats:sub> was only 2\u20133 times higher than for CD<jats:sub>2</jats:sub>=H<jats:sub>4</jats:sub>MPT dehydrogenation to CD\u2261H<jats:sub>4</jats:sub>MPT<jats:sup>+</jats:sup> and HD. Such a small primary isotope effect indicates that the breakage of the C\u2010H bond in the methylene group of CH<jats:sub>2</jats:sub>=H<jats:sub>4</jats:sub>MPT was only rate\u2010limiting when hydrogen was substituted by a deuterium.</jats:p>",
  "DOI": "10.1111/j.1432-1033.1995.372_1.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2004,
        9,
        8
      ]
    ],
    "date-time": "2004-09-08T08:04:27Z",
    "timestamp": 1094630667000
  },
  "page": "372-376",
  "source": "Crossref",
  "is-referenced-by-count": 30,
  "title": "Hydrogen Isotope Effects in the Reactions Catalyzed by H<sub>2</sub>\u2010forming <i>N</i><sup>5</sup>,<i>N</i><sup>10</sup>\u2010Methylenetetrahydromethanopterin Dehydrogenase from Methanogenic Archaea",
  "prefix": "10.1111",
  "volume": "233",
  "author": [
    {
      "given": "Andreas R.",
      "family": "Klein",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Gudrun C.",
      "family": "Hartmann",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Rudolf K.",
      "family": "Thauer",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2004,
        9,
        8
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_3_2_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0005-2728(94)90036-1"
    },
    {
      "key": "e_1_2_3_3_1",
      "doi-asserted-by": "crossref",
      "DOI": "10.1002/ange.19951072011",
      "article-title": "Zum Katalysemechanismus der Nickel\u2010 und Eisen\u2010Cluster\u2010freien Hydrogenase aus methanogenen Archaea: Enzymatische H2\u2010Aktivierung ohne Metall und ihre Analogie zur Chemie der Alkane in supersaurer L\u00f6sung",
      "author": "Berkessel A.",
      "year": "1995",
      "journal-title": "Angew. Chem."
    },
    {
      "key": "e_1_2_3_4_1",
      "first-page": "634",
      "article-title": "Chemistry and spectroscopy in strongly acidic solutions: Reversible reaction between alipathic carbonium ions and hydrogen",
      "volume": "67",
      "author": "Bickel A. F.",
      "year": "1967",
      "journal-title": "J. Chem. Soc., Chem. Commun."
    },
    {
      "key": "e_1_2_3_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1992.tb17502.x"
    },
    {
      "key": "e_1_2_3_6_1",
      "first-page": "1",
      "volume-title": "The enzymes",
      "author": "Cleland W. W.",
      "year": "1970"
    },
    {
      "key": "e_1_2_3_7_1",
      "volume-title": "Enzyme mechanism from isotope effects",
      "author": "Cook P. F.",
      "year": "1991"
    },
    {
      "key": "e_1_2_3_8_1",
      "first-page": "203",
      "volume-title": "Enzyme mechanism from isotope effects",
      "author": "Cook P. F.",
      "year": "1991"
    },
    {
      "key": "e_1_2_3_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/ac60260a013"
    },
    {
      "key": "e_1_2_3_10_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(17)42721-9"
    },
    {
      "key": "e_1_2_3_11_1",
      "unstructured": "Hartmann G. C.(1994)H2\u2010bildendeN5 N10\u2010Methylentetrahydromethanopterin\u2010Dehydrogenase und andere Enzyme der Methanogenese ausMethanococcus thermolithotrophicus Diploma Thesis Philipps\u2010Universit\u00e4t Marburg."
    },
    {
      "key": "e_1_2_3_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.143.2.628-636.1980"
    },
    {
      "key": "e_1_2_3_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00418186"
    },
    {
      "key": "e_1_2_3_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00644273"
    },
    {
      "key": "e_1_2_3_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jb.177.9.2460-2468.1995"
    },
    {
      "key": "e_1_2_3_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/ange.19931050604"
    },
    {
      "key": "e_1_2_3_17_1",
      "first-page": "74",
      "volume-title": "Enzyme mechanism from isotope effects",
      "author": "Quinn D. M.",
      "year": "1991"
    },
    {
      "key": "e_1_2_3_18_1",
      "first-page": "321",
      "volume-title": "The enzymes",
      "author": "Richards J. H.",
      "year": "1970"
    },
    {
      "key": "e_1_2_3_19_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0014-5793(92)81522-N"
    },
    {
      "key": "e_1_2_3_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00179a027"
    },
    {
      "key": "e_1_2_3_21_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/ja00115a035"
    },
    {
      "key": "e_1_2_3_22_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00244962"
    },
    {
      "key": "e_1_2_3_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1993.tb17657.x"
    },
    {
      "key": "e_1_2_3_24_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/978-1-4615-2391-8_5"
    },
    {
      "key": "e_1_2_3_25_1",
      "unstructured": "Vaupel M.(1993)Untersuchungen mitMethanobacterium thermoformicicumundMethanobacterium formicicumzur Rolle von H2\u2010bildenderN5 N10\u2010Methylentetrahydromethanopterin Dehydrogenase in methanogenen Archaea. Diploma Thesis Philipps\u2010Universit\u00e4t Marburg."
    },
    {
      "key": "e_1_2_3_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1991.tb16491.x"
    },
    {
      "key": "e_1_2_3_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(93)90570-G"
    },
    {
      "key": "e_1_2_3_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0014-5793(90)80649-4"
    },
    {
      "key": "e_1_2_3_29_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1432-1033.1992.tb17215.x"
    }
  ],
  "container-title": "European Journal of Biochemistry",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1432-1033.1995.372_1.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1995.372_1.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        31
      ]
    ],
    "date-time": "2023-10-31T12:56:57Z",
    "timestamp": 1698757017000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://febs.onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1995.372_1.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1995,
        10
      ]
    ]
  },
  "references-count": 28,
  "journal-issue": {
    "issue": "1",
    "published-print": {
      "date-parts": [
        [
          1995,
          10
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1111/j.1432-1033.1995.372_1.x"
  ],
  "URL": "http://dx.doi.org/10.1111/j.1432-1033.1995.372_1.x",
  "relation": {},
  "ISSN": [
    "0014-2956",
    "1432-1033"
  ],
  "subject": [],
  "container-title-short": "European Journal of Biochemistry",
  "published": {
    "date-parts": [
      [
        1995,
        10
      ]
    ]
  }
}