Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.jmb.2008.02.041
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dc.titleCrystal Structure of the Polyextremophilic α-Amylase AmyB from Halothermothrix orenii: Details of a Productive Enzyme-Substrate Complex and an N Domain with a Role in Binding Raw Starch
dc.contributor.authorTan, T.-C.
dc.contributor.authorMijts, B.N.
dc.contributor.authorSwaminathan, K.
dc.contributor.authorPatel, B.K.C.
dc.contributor.authorDivne, C.
dc.date.accessioned2014-10-27T08:25:03Z
dc.date.available2014-10-27T08:25:03Z
dc.date.issued2008-05-09
dc.identifier.citationTan, T.-C., Mijts, B.N., Swaminathan, K., Patel, B.K.C., Divne, C. (2008-05-09). Crystal Structure of the Polyextremophilic α-Amylase AmyB from Halothermothrix orenii: Details of a Productive Enzyme-Substrate Complex and an N Domain with a Role in Binding Raw Starch. Journal of Molecular Biology 378 (4) : 850-868. ScholarBank@NUS Repository. https://doi.org/10.1016/j.jmb.2008.02.041
dc.identifier.issn00222836
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/100369
dc.description.abstractThe gene for a membrane-bound, halophilic, and thermostable α-amylase, AmyB, from Halothermothrix orenii was cloned and sequenced. The crystal structure shows that, in addition to the typical domain organization of family 13 glycoside hydrolases, AmyB carries an additional N-terminal domain (N domain) that forms a large groove-the N-C groove-some 30 Å away from the active site. The structure of AmyB with the inhibitor acarbose at 1.35 Å resolution shows that a nonasaccharide has been synthesized through successive transglycosylation reactions of acarbose. Unexpectedly, in a complex of wild-type AmyB with α-cyclodextrin and maltoheptaose at 2.2 Å resolution, a maltotetraose molecule is bound in subsites - 1 to + 3, spanning the cleavage point at - 1/+ 1, with the - 1 glucosyl residue present as a 2So skew boat. This wild-type AmyB complex was obtained in the presence of a large excess of substrate, a condition under which it is possible to capture Michaelis complexes, which may explain the observed binding across - 1/+ 1 and ring distortion. We observe three methionine side chains that serve as "binding platforms" for glucosyl rings in AmyB, a seemingly rare occurrence in carbohydrate-binding proteins. The structures and results from the biochemical characterization of AmyB and AmyB lacking the N domain show that the N domain increases binding of the enzyme to raw starch. Furthermore, theoretical modeling suggests that the N-C groove can accommodate, spatially and chemically, large substrates such as A-starch. © 2008 Elsevier Ltd. All rights reserved.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1016/j.jmb.2008.02.041
dc.sourceScopus
dc.subjectα-amylase
dc.subjectenzyme-substrate complex
dc.subjectmethionine interaction
dc.subjectN domain
dc.subjectpolyextremophilic
dc.typeArticle
dc.contributor.departmentBIOLOGICAL SCIENCES
dc.description.doi10.1016/j.jmb.2008.02.041
dc.description.sourcetitleJournal of Molecular Biology
dc.description.volume378
dc.description.issue4
dc.description.page850-868
dc.description.codenJMOBA
dc.identifier.isiut000256180100008
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