3AUJ

Structure of diol dehydratase complexed with glycerol

Summary for 3AUJ

• Entry DOI: 10.2210/pdb3auj/pdb
• Related: 1DIO 1EEX 1EGM 1EGV 1IWB 1UC4 1UC5
• Descriptor: Diol dehydrase alpha subunit, Diol dehydrase beta subunit, Diol dehydrase gamma subunit, ... (8 entities in total)
• Functional Keywords: alpha/beta barrel, lyase
• Biological source: Klebsiella oxytoca; More
• Total number of polymer chains: 6
• Total formula weight: 210991.89

Authors

Yamanishi, M.,Kinoshita, K.,Fukuoka, M.,Shibata, T.,Tobimatsu, T.,Toraya, T. (deposition date: 2011-02-07, release date: 2012-02-22, Last modification date: 2023-11-01)

Primary citation

Yamanishi, M.,Kinoshita, K.,Fukuoka, M.,Saito, T.,Tanokuchi, A.,Ikeda, Y.,Obayashi, H.,Mori, K.,Shibata, N.,Tobimatsu, T.,Toraya, T.
Redesign of coenzyme B(12) dependent diol dehydratase to be resistant to the mechanism-based inactivation by glycerol and act on longer chain 1,2-diols
Febs J., 279:793-804, 2012

PubMed Abstract: Coenzyme B(12) dependent diol dehydratase undergoes mechanism-based inactivation by glycerol, accompanying the irreversible cleavage of the coenzyme Co-C bond. Bachovchin et al. [Biochemistry16, 1082-1092 (1977)] reported that glycerol bound in the G(S) conformation, in which the pro-S-CH(2) OH group is oriented to the hydrogen-abstracting site, primarily contributes to the inactivation reaction. To understand the mechanism of inactivation by glycerol, we analyzed the X-ray structure of diol dehydratase complexed with cyanocobalamin and glycerol. Glycerol is bound to the active site preferentially in the same conformation as that of (S)-1,2-propanediol, i.e. in the G(S) conformation, with its 3-OH group hydrogen bonded to Serα301, but not to nearby Glnα336. k(inact) of the Sα301A, Qα336A and Sα301A/Qα336A mutants with glycerol was much smaller than that of the wild-type enzyme. k(cat) /k(inact) showed that the Sα301A and Qα336A mutants are substantially more resistant to glycerol inactivation than the wild-type enzyme, suggesting that Serα301 and Glnα336 are directly or indirectly involved in the inactivation. The degree of preference for (S)-1,2-propanediol decreased on these mutations. The substrate activities towards longer chain 1,2-diols significantly increased on the Sα301A/Qα336A double mutation, probably because these amino acid substitutions yield more space for accommodating a longer alkyl group on C3 of 1,2-diols. Database Structural data are available in the Protein Data Bank under the accession number 3AUJ. Structured digital abstract • Diol dehydrase gamma subunit, Diol dehydrase beta subunit and Diol dehydrase alpha subunit physically interact by X-ray crystallography (View interaction).

PubMed: 22221669
DOI: 10.1111/j.1742-4658.2012.08470.x

Experimental method

X-RAY DIFFRACTION (2.1 Å)