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PMID: 8099449 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular and active-site structure of a Bacillus 1,3-1,4-beta-glucanase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 11 ·1993-06-01 ·Pages 5287-91

Keitel T, Simon O, Borriss R, Heinemann U

Abstract

The three-dimensional structure of the hybrid Bacillus 1,3-1,4-beta-glucanase (beta-glucanase; 1,3-1,4-beta-D-glucan 4-glucanohydrolase, lichenase, EC 3.2.1.73) designated H(A16-M) was determined by x-ray crystallography at a resolution of 2.0 A and refined to an R value of 16.4% using stereochemical restraints. The protein molecule consists mainly of two seven-stranded antiparallel beta-pleated sheets arranged atop each other to form a compact, sandwich-like structure. A channel crossing one side of the protein molecule accommodates an inhibitor, 3,4-epoxybutyl beta-D-cellobioside, which binds covalently to the side chain of Glu-105, as seen in a crystal structure analysis at 2.8-A resolution of the protein-inhibitor complex (R = 16.8%). That Glu-105 may be indispensible for enzyme catalysis by H(A16-M) is suggested by site-directed mutagenesis of this residue, which inevitably leads to an inactive enzyme.

MeSH Terms
Amino Acid Sequence Bacillus/enzymology Binding Sites Carbohydrate Sequence Epoxy Compounds/metabolism,pharmacology Glucans/metabolism Glucosides/metabolism,pharmacology Glutamates Glutamic Acid Glycoside Hydrolases/chemistry,genetics,metabolism Hydrogen Bonding Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Structure, Secondary Recombinant Proteins/chemistry,metabolism Substrate Specificity X-Ray Diffraction
Chemicals
Epoxy Compounds Glucans Glucosides Glutamates Recombinant Proteins 3,4-epoxybutyl-beta-cellobioside Glutamic Acid Glycoside Hydrolases licheninase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Keitel T
Institut für Kristallographie, Freie Universität, Berlin, Federal Republic of Germany.
Simon O
Borriss R
Heinemann U
References (29)
29 references, click to expand
  1. Three-dimensional structure of cellobiohydrolase II from Trichoderma reesei.
    Science. 1990 Jul 27;249(4967):380-6 PMID: 2377893
  2. Active site-directed inhibition by optically pure epoxyalkyl cellobiosides reveals differences in active site geometry of two 1,3-1,4-beta-D-glucan 4-glucanohydrolases. The importance of epoxide stereochemistry for enzyme inactivation.
    J Biol Chem. 1991 Jun 25;266(18):11628-31 PMID: 1904865
  3. Molecular cloning, expression and nucleotide sequence of the endo-beta-1,3-1,4-D-glucanase gene from Bacillus licheniformis. Predictive structural analyses of the encoded polypeptide.
    Eur J Biochem. 1991 Apr 23;197(2):337-43 PMID: 2026156
  4. Two beta-glycanase genes are clustered in Bacillus polymyxa: molecular cloning, expression, and sequence analysis of genes encoding a xylanase and an endo-beta-(1,3)-(1,4)-glucanase.
    J Bacteriol. 1991 Dec;173(23):7705-10 PMID: 1938968
  5. A new substrate for investigating the specificity of beta-glucan hydrolases.
    FEBS Lett. 1975 Apr 1;52(2):202-7 PMID: 236912
  6. DNA sequence of a Fibrobacter succinogenes mixed-linkage beta-glucanase (1,3-1,4-beta-D-glucan 4-glucanohydrolase) gene.
    J Bacteriol. 1990 Jul;172(7):3837-41 PMID: 2193918
  7. Refinement of human lysozyme at 1.5 A resolution analysis of non-bonded and hydrogen-bond interactions.
    J Mol Biol. 1981 Nov 15;152(4):737-62 PMID: 7334520
  8. Three dimensional structure of porcine pancreatic alpha-amylase at 2.9 A resolution. Role of calcium in structure and activity.
    EMBO J. 1987 Dec 20;6(13):3909-16 PMID: 3502087
  9. Structure of the beta-1,3-1,4-glucanase gene of Bacillus macerans: homologies to other beta-glucanases.
    Mol Gen Genet. 1990 Jul;222(2-3):278-83 PMID: 2274030
  10. Double stranded DNA sequencing as a choice for DNA sequencing.
    Nucleic Acids Res. 1988 Feb 11;16(3):1220 PMID: 3344219
  11. Carboxyl-carboxylate interactions in proteins.
    Nature. 1982 Jan 7;295(5844):79-80 PMID: 7057876
  12. Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features.
    Biopolymers. 1983 Dec;22(12):2577-637 PMID: 6667333
  13. The beta-glucanase gene from Bacillus amyloliquefaciens shows extensive homology with that of Bacillus subtilis.
    Gene. 1986;49(2):177-87 PMID: 3106158
  14. Determination of the three-dimensional solution structure of the C-terminal domain of cellobiohydrolase I from Trichoderma reesei. A study using nuclear magnetic resonance and hybrid distance geometry-dynamical simulated annealing.
    Biochemistry. 1989 Sep 5;28(18):7241-57 PMID: 2554967
  15. Structure and possible catalytic residues of Taka-amylase A.
    J Biochem. 1984 Mar;95(3):697-702 PMID: 6609921
  16. Resolution of phase ambiguity in macromolecular crystallography.
    Methods Enzymol. 1985;115:90-112 PMID: 4079800
  17. Hybrid bacillus endo-(1-3,1-4)-beta-glucanases: construction of recombinant genes and molecular properties of the gene products.
    Carlsberg Res Commun. 1989;54(2):41-54 PMID: 2673278
  18. Crystallization of the hybrid Bacillus (1-3,1-4)-beta-glucanase H(A16-M).
    J Mol Biol. 1991 Apr 20;218(4):703-4 PMID: 2023245
  19. Hybrid Bacillus (1-3,1-4)-beta-glucanases: engineering thermostable enzymes by construction of hybrid genes.
    Mol Gen Genet. 1991 Feb;225(2):177-85 PMID: 2005860
  20. Crystallographic determination of the mode of binding of oligosaccharides to T4 bacteriophage lysozyme: implications for the mechanism of catalysis.
    J Mol Biol. 1981 Apr 25;147(4):523-43 PMID: 7277499
  21. The DNA sequence of the gene and genetic control sites for the excreted B. subtilis enzyme beta-glucanase.
    Nucleic Acids Res. 1984 Jul 11;12(13):5355-67 PMID: 6087283
  22. Essential catalytic role of Glu134 in endo-beta-1,3-1,4-D-glucan 4-glucanohydrolase from B. licheniformis as determined by site-directed mutagenesis.
    FEBS Lett. 1992 Aug 17;308(2):141-5 PMID: 1354172
  23. The crystal structure of pea lectin at 3.0-A resolution.
    J Biol Chem. 1986 Dec 15;261(35):16518-27 PMID: 3782132
  24. Differences in active site structure in a family of beta-glucan endohydrolases deduced from the kinetics of inactivation by epoxyalkyl beta-oligoglucosides.
    J Biol Chem. 1989 Mar 25;264(9):4939-47 PMID: 2494179
  25. Crystallographic R factor refinement by molecular dynamics.
    Science. 1987 Jan 23;235(4787):458-60 PMID: 17810339
  26. Primary structure of the (1-->3,1-->4)-beta-D-glucan 4-glucohydrolase from barley aleurone.
    Proc Natl Acad Sci U S A. 1986 Apr;83(7):2081-5 PMID: 16593676
  27. Stereochemically restrained refinement of macromolecular structures.
    Methods Enzymol. 1985;115:252-70 PMID: 3841182
  28. Identification of glutamic acid 105 at the active site of Bacillus amyloliquefaciens 1,3-1,4-beta-D-glucan 4-glucanohydrolase using epoxide-based inhibitors.
    J Biol Chem. 1992 Dec 15;267(35):25059-66 PMID: 1360982
  29. Structure of the Clostridium thermocellum gene licB and the encoded beta-1,3-1,4-glucanase. A catalytic region homologous to Bacillus lichenases joined to the reiterated domain of clostridial cellulases.
    Eur J Biochem. 1992 Feb 15;204(1):13-9 PMID: 1740123
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.11.5287
Published
1993-06-01
Pages
5287-91
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC46701
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.90.11.5287 PMC Full Text View journal details
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