@article{6c4de02744824498b5c1510c323ad500,
title = "Physical association of the catalytic and helper modules of a family-9 glycoside hydrolase is essential for activity",
abstract = "Clostridium thermocellum cellulase 9I (Cel9I) is a non-cellulosomal tri-modular enzyme, consisting of a family-9 glycoside hydrolase (GH9) catalytic module and two family-3 carbohydrate-binding modules (CBM3c and CBM3b). The presence of CBM3c was previously shown to be essential for activity, however the mechanism by which it functions is unclear. We expressed the three recombinant modules independently in Escherichia coli and examined their interactions. Non-denaturing gel electrophoresis, isothermal titration calorimetry, and affinity purification of the GH9-CBM3c complex revealed a specific non-covalent binding interaction between the GH9 module and CBM3c. Their physical association was shown to recover 60-70% of the intact Cel9I endoglucanase activity. Structured summary: MINT-6946626:. Cel9I (uniprotkb:Q02934) and Cel9I (uniprotkb:Q02934) bind (MI:0407) by comigration in non-denaturing gel electrophoresis (MI:0404). MINT-6946649:. Cel9I (uniprotkb:Q02934) and Cel9I (uniprotkb:Q02934) bind (MI:0407) by molecular sieving (MI:0071). MINT-6946687:. Cel9I (uniprotkb:Q02934) and Cel9I (uniprotkb:Q02934) bind (MI:0407) by isothermal titration calorimetry (MI:0065). MINT-6946706:. Cel9I (uniprotkb:Q02934) binds (MI:0407) to Cel9I (uniprotkb:Q02934) by pull down (MI:0096).",
keywords = "Cellulase, Cellulose, Endoglucanase, Family-3 carbohydrate-binding module (CBM3), Family-9 glycoside hydrolases (GH9)",
author = "Tal Burstein and Michal Shulman and Sadanari Jindou and Svetlana Petkun and Felix Frolow and Yuval Shoham and Bayer, {Edward A.} and Raphael Lamed",
note = "Funding Information: The authors are grateful to Oshik Segev (Tel Aviv University) for his assistance with the ITC data. This research was supported by the Israel Science Foundation (Grant Nos. 422/05, 159/07 and 291/08), and by grants from the United States–Israel Binational Science Foundation (BSF), Jerusalem, Israel. Y.S. holds the Erwin and Rosl Pollak Chair in Biotechnology at the Technion, and E.A.B. is the incumbent of The Maynard I. and Elaine Wishner Chair of Bio-organic Chemistry.",
year = "2009",
month = mar,
day = "4",
doi = "10.1016/j.febslet.2009.02.013",
language = "אנגלית",
volume = "583",
pages = "879--884",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "John Wiley and Sons Inc.",
number = "5",
}