Characterization of Choline Trimethylamine-Lyase Expands the Chemistry of Glycyl Radical Enzymes

被引：112

作者：

Craciun, Smaranda ^{[1
]}

Marks, Jonathan A. ^{[1
]}

Balskus, Emily P. ^{[1
]}

机构：

[1] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA

来源：

ACS CHEMICAL BIOLOGY | 2014年 / 9卷 / 07期

关键词：

PYRUVATE FORMATE-LYASE; ETHANOLAMINE AMMONIA-LYASE; CRYSTAL-STRUCTURES; ESCHERICHIA-COLI; DEGRADATION; MECHANISM;

D O I：

10.1021/cb500113p

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The recently identified glycyl radical enzyme (GRE) homologue choline trimethylamine-lyase (CutC) participates in the anaerobic conversion of choline to trimethylamine (TMA), a widely distributed microbial metabolic transformation that occurs in the human gut and is linked to disease. The proposed biochemical function of CutC, C-N bond cleavage, represents new reactivity for the GRE family. Here we describe the in vitro characterization of CutC and its activating protein CutD. We have observed CutD-mediated formation of a glycyl radical on CutC using EPR spectroscopy and have demonstrated that activated CutC processes choline to trimethylamine and acetaldehyde. Surveys of potential alternate CutC substrates uncovered a strict specificity for choline. Homology modeling and mutagenesis experiments revealed essential CutC active site residues. Overall, this work establishes that CutC is a GRE of unique function and a molecular marker for anaerobic choline metabolism.

引用

页码：1408 / 1413

页数：6

共 33 条

[1] Purification and some properties of wild-type and N-terminal-truncated ethanolamine ammonia-lyase of Escherichia coli [J].

Akita, Keita ;

Hieda, Naoki ;

Baba, Nobuyuki ;

Kawaguchi, Satoshi ;

Sakamoto, Hirohisa ;

Nakanishi, Yuka ;

Yamanishi, Mamoru ;

Mori, Koichi ;

Toraya, Tetsuo .

JOURNAL OF BIOCHEMISTRY, 2010, 147 (01) :83-93

[2] Microbial conversion of choline to trimethylamine requires a glycyl radical enzyme [J].

Craciun, Smaranda ;

Balskus, Emily P. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2012, 109 (52) :21307-21312

[3] Pyruvate Formate-lyase and Its Activation by Pyruvate Formate-lyase Activating Enzyme [J].

Crain, Adam V. ;

Broderick, Joan B. .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2014, 289 (09) :5723-5729

[4] Metabolic profiling reveals a contribution of gut microbiota to fatty liver phenotype in insulin-resistant mice [J].

Dumas, Marc-Emmanuel ;

Barton, Richard H. ;

Toye, Ayo ;

Cloarec, Olivier ;

Blancher, Christine ;

Rothwell, Alice ;

Fearnside, Jane ;

Tatoud, Roger ;

Blanc, Veronique ;

Lindon, John C. ;

Mitchell, Steve C. ;

Holmes, Elaine ;

McCarthy, Mark I. ;

Scott, James ;

Gauguier, Dominique ;

Nicholson, Jeremy K. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (33) :12511-12516

[5] Glycerol Dehydratation by the B12-Independent Enzyme May Not Involve the Migration of a Hydroxyl Group: A Computational Study [J].

Feliks, Mikolaj ;

Ullmann, G. Matthias .

JOURNAL OF PHYSICAL CHEMISTRY B, 2012, 116 (24) :7076-7087

[6] Radical mechanisms of enzymatic catalysis [J].

Frey, PA .

ANNUAL REVIEW OF BIOCHEMISTRY, 2001, 70 :121-148

[7]

HAYWARD HR, 1960, J BIOL CHEM, V235, P538

[8] UTILIZATION OF TRIMETHYLAMINE AND OTHER N-METHYL COMPOUNDS FOR GROWTH AND METHANE FORMATION BY METHANOSARCINA-BARKERI [J].

HIPPE, H ;

CASPARI, D ;

FIEBIG, K ;

GOTTSCHALK, G .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (01) :494-498

[9] Using comparative genomics to uncover new kinds of protein-based metabolic organelles in bacteria [J].

Jorda, Julien ;

Lopez, David ;

Wheatley, Nicole M. ;

Yeates, Todd O. .

PROTEIN SCIENCE, 2013, 22 (02) :179-195

[10] Structure, function, and mechanism of ribonucleotide reductases [J].

Kolberg, M ;

Strand, KR ;

Graff, P ;

Andersson, KK .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 2004, 1699 (1-2) :1-34

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