Structure, computational and biochemical analysis of PcCel45A endoglucanase from Phanerochaete chrysosporium and catalytic mechanisms of GH45 subfamily C members

Research output: Contribution to journalJournal article – Annual report year: 2018Researchpeer-review

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Structure, computational and biochemical analysis of PcCel45A endoglucanase from Phanerochaete chrysosporium and catalytic mechanisms of GH45 subfamily C members. / Godoy, Andre S.; Pereira, Caroline S.; Ramia, Marina Paglione; Silveira, Rodrigo L.; Camilo, Cesar M.; Kadowaki, Marco A.; Lange, Lene; Busk, Peter K.; Nascimento, Alessandro S.; Skaf, Munir S.; Polikarpov, Igor.

In: Scientific Reports, Vol. 8, 3678, 2018.

Research output: Contribution to journalJournal article – Annual report year: 2018Researchpeer-review

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Godoy, Andre S. ; Pereira, Caroline S. ; Ramia, Marina Paglione ; Silveira, Rodrigo L. ; Camilo, Cesar M. ; Kadowaki, Marco A. ; Lange, Lene ; Busk, Peter K. ; Nascimento, Alessandro S. ; Skaf, Munir S. ; Polikarpov, Igor. / Structure, computational and biochemical analysis of PcCel45A endoglucanase from Phanerochaete chrysosporium and catalytic mechanisms of GH45 subfamily C members. In: Scientific Reports. 2018 ; Vol. 8.

Bibtex

@article{c60641e80d364353aa155388a0aaa845,
title = "Structure, computational and biochemical analysis of PcCel45A endoglucanase from Phanerochaete chrysosporium and catalytic mechanisms of GH45 subfamily C members",
abstract = "The glycoside hydrolase family 45 (GH45) of carbohydrate modifying enzymes is mostly comprised of {\ss}-1,4-endoglucanases. Significant diversity between the GH45 members has prompted the division of this family into three subfamilies: A, B and C, which may differ in terms of the mechanism, general architecture, substrate binding and cleavage. Here, we use a combination of X-ray crystallography, bioinformatics, enzymatic assays, molecular dynamics simulations and site-directed mutagenesis experiments to characterize the structure, substrate binding and enzymatic specificity of the GH45 subfamily C endoglucanase from Phanerochaete chrysosporium (PcCel45A). We investigated the role played by different residues in the binding of the enzyme to cellulose oligomers of different lengths and examined the structural characteristics and dynamics of PcCel45A that make subfamily C so dissimilar to other members of the GH45 family. Due to the structural similarity shared between PcCel45A and domain I of expansins, comparative analysis of their substrate binding was also carried out. Our bioinformatics sequence analyses revealed that the hydrolysis mechanisms in GH45 subfamily C is not restricted to use of the imidic asparagine as a general base in the {"}Newton's cradle{"} catalytic mechanism recently proposed for this subfamily.",
author = "Godoy, {Andre S.} and Pereira, {Caroline S.} and Ramia, {Marina Paglione} and Silveira, {Rodrigo L.} and Camilo, {Cesar M.} and Kadowaki, {Marco A.} and Lene Lange and Busk, {Peter K.} and Nascimento, {Alessandro S.} and Skaf, {Munir S.} and Igor Polikarpov",
year = "2018",
doi = "10.1038/s41598-018-21798-9",
language = "English",
volume = "8",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

RIS

TY - JOUR

T1 - Structure, computational and biochemical analysis of PcCel45A endoglucanase from Phanerochaete chrysosporium and catalytic mechanisms of GH45 subfamily C members

AU - Godoy, Andre S.

AU - Pereira, Caroline S.

AU - Ramia, Marina Paglione

AU - Silveira, Rodrigo L.

AU - Camilo, Cesar M.

AU - Kadowaki, Marco A.

AU - Lange, Lene

AU - Busk, Peter K.

AU - Nascimento, Alessandro S.

AU - Skaf, Munir S.

AU - Polikarpov, Igor

PY - 2018

Y1 - 2018

N2 - The glycoside hydrolase family 45 (GH45) of carbohydrate modifying enzymes is mostly comprised of ß-1,4-endoglucanases. Significant diversity between the GH45 members has prompted the division of this family into three subfamilies: A, B and C, which may differ in terms of the mechanism, general architecture, substrate binding and cleavage. Here, we use a combination of X-ray crystallography, bioinformatics, enzymatic assays, molecular dynamics simulations and site-directed mutagenesis experiments to characterize the structure, substrate binding and enzymatic specificity of the GH45 subfamily C endoglucanase from Phanerochaete chrysosporium (PcCel45A). We investigated the role played by different residues in the binding of the enzyme to cellulose oligomers of different lengths and examined the structural characteristics and dynamics of PcCel45A that make subfamily C so dissimilar to other members of the GH45 family. Due to the structural similarity shared between PcCel45A and domain I of expansins, comparative analysis of their substrate binding was also carried out. Our bioinformatics sequence analyses revealed that the hydrolysis mechanisms in GH45 subfamily C is not restricted to use of the imidic asparagine as a general base in the "Newton's cradle" catalytic mechanism recently proposed for this subfamily.

AB - The glycoside hydrolase family 45 (GH45) of carbohydrate modifying enzymes is mostly comprised of ß-1,4-endoglucanases. Significant diversity between the GH45 members has prompted the division of this family into three subfamilies: A, B and C, which may differ in terms of the mechanism, general architecture, substrate binding and cleavage. Here, we use a combination of X-ray crystallography, bioinformatics, enzymatic assays, molecular dynamics simulations and site-directed mutagenesis experiments to characterize the structure, substrate binding and enzymatic specificity of the GH45 subfamily C endoglucanase from Phanerochaete chrysosporium (PcCel45A). We investigated the role played by different residues in the binding of the enzyme to cellulose oligomers of different lengths and examined the structural characteristics and dynamics of PcCel45A that make subfamily C so dissimilar to other members of the GH45 family. Due to the structural similarity shared between PcCel45A and domain I of expansins, comparative analysis of their substrate binding was also carried out. Our bioinformatics sequence analyses revealed that the hydrolysis mechanisms in GH45 subfamily C is not restricted to use of the imidic asparagine as a general base in the "Newton's cradle" catalytic mechanism recently proposed for this subfamily.

U2 - 10.1038/s41598-018-21798-9

DO - 10.1038/s41598-018-21798-9

M3 - Journal article

VL - 8

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 3678

ER -