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430 a.a.
Key reference
DOI no: 10.1111/j.1432-1033.2004.04409.x Eur J Biochem 271:4495-4506 (2004) PubMed id: 15560790
Three-dimensional structure of a thermostable native cellobiohydrolase, CBH IB, and molecular characterization of the cel7 gene from the filamentous fungus, Talaromyces emersonii. A.Grassick, P.G.Murray, R.Thompson, C.M.Collins, L.Byrnes, G.Birrane, T.M.Higgins, M.G.Tuohy. ABSTRACT
The X-ray structure of native cellobiohydrolase IB (CBH IB) from the filamentous fungus Talaromyces emersonii, PDB 1Q9H, was solved to 2.4 A by molecular replacement. 1Q9H is a glycoprotein that consists of a large, single domain with dimensions of approximately 60 A x 40 A x 50 A and an overall beta-sandwich structure, the characteristic fold of Family 7 glycosyl hydrolases (GH7). It is the first structure of a native glycoprotein and cellulase from this thermophilic eukaryote. The long cellulose-binding tunnel seen in GH7 Cel7A from Trichoderma reesei is conserved in 1Q9H, as are the catalytic residues. As a result of deletions and other changes in loop regions, the binding and catalytic properties of T. emersonii 1Q9H are different. The gene (cel7) encoding CBH IB was isolated from T. emersonii and expressed heterologously with an N-terminal polyHis-tag, in Escherichia coli. The deduced amino acid sequence of cel7 is homologous to fungal cellobiohydrolases in GH7. The recombinant cellobiohydrolase was virtually inactive against methylumberiferyl-cellobioside and chloronitrophenyl-lactoside, but partial activity could be restored after refolding of the urea-denatured enzyme. Profiles of cel7 expression in T. emersonii, investigated by Northern blot analysis, revealed that expression is regulated at the transcriptional level. Putative regulatory element consensus sequences for cellulase transcription factors have been identified in the upstream region of the cel7 genomic sequence.
Selected figure(s)
Figure 3.
Fig. 3. Electron maps at the two N-glycosylation sites. (A) Asn267 with two GlcNAc (2-amino-2-N-acetylamino-D-glucose) residues. (B) Asn431 with one GlcNAc residue.Figure 7.
Fig. 7. C-alpha trace of 1Q9H(yellow) superimposed on the C-alpha trace of 5 Cel(white), illustrating the more open active site of 1Q9H. The sugar residues are superimposed in blue. The catalytic residues are shown in red. The figure was drawn by using TURBO.The above figures are reprinted by permission from the Federation of European Biochemical Societies: Eur J Biochem (2004, 271, 4495-4506) copyright 2004. Figures were selected by an automated process.
Literature references that cite this PDB file's key reference
PubMed id Reference
19189377 B.Mertz, X.Gu, and P.J.Reilly (2009).
Analysis of functional divergence within two structurally related glycoside hydrolase families.Biopolymers, 91, 478-495. 19774110 M.Dashtban, H.Schraft, and W.Qin (2009).
Fungal bioconversion of lignocellulosic residues; opportunities & perspectives.Int J Biol Sci, 5, 578-595.
19148633 S.P.Voutilainen, H.Boer, M.Alapuranen, J.Jänis, J.Vehmaanperä, and A.Koivula (2009).
Improving the thermostability and activity of Melanocarpus albomyces cellobiohydrolase Cel7B.Appl Microbiol Biotechnol, 83, 261-272.
18687686 A.J.O'Donoghue, C.S.Mahon, D.H.Goetz, J.M.O'Malley, D.M.Gallagher, M.Zhou, P.G.Murray, C.S.Craik, and M.G.Tuohy (2008).
Inhibition of a secreted glutamic peptidase prevents growth of the fungus Talaromyces emersonii.J Biol Chem, 283, 29186-29195.
18499583 T.Parkkinen, A.Koivula, J.Vehmaanperä, and J.Rouvinen (2008).
Crystal structures of Melanocarpus albomyces cellobiohydrolase Cel7B in complex with cello-oligomers show high flexibility in the substrate binding.Protein Sci, 17, 1383-1394.
PDB codes: 2rfw 2rfy 2rfz 2rg0 17768346 T.Parkkinen, A.Koivula, J.Vehmaanperä, and J.Rouvinen (2007).
Preliminary X-ray analysis of cellobiohydrolase Cel7B from Melanocarpus albomyces.Acta Crystallogr Sect F Struct Biol Cryst Commun, 63, 754-757.
17277884 Y.Hou, T.Wang, H.Long, and H.Zhu (2007).
Cloning, sequencing and expression analysis of the first cellulase gene encoding cellobiohydrolase 1 from a cold-adaptive Penicillium chrysogenum FS010.Acta Biochim Biophys Sin (Shanghai), 39, 101-107. The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.
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