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Probing promiscuity

structural studies of Phanerochaete chrysosporium Laminarinase 16A

Vasur, Jonas (2009). Probing promiscuity. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis agriculturae Sueciae, 1652-6880 ; 2009:98
ISBN 978-91-576-7445-6
[Doctoral thesis]

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Abstract

The glycoside hydrolase Laminarinase 16A (Lam16A) from the white-rot fungus Phanerochaete chrysosporiumı is able to bind and cleave two different substrates in a specific manner. It has been called "non-specific", but we find that binding promiscuity and catalytic promiscuity better define this phenomenon. We have published high-resolution structures of wildtype Lam16A alone (Paper I) and in complex with oligosaccharide products from hydrolysis of laminarin and lichenan (up to 1.1 Å resolution) (Paper II) as well as structures of the catalytic nucleophile mutant Lam16A E115S in complex with laminariheptaose and alpha-laminariheptaosyl fluoride (Paper III, submitted). A beta-sandwich forms the substrate binding cleft, evenly open at both ends. The Lam16A ligand/complex structures, especially the catalytic mutants (Paper IV, manuscript), illustrate why two beta-1,3-linked glucose residues are required at the -2 and -1 subsites and why no beta-1,6-branch can be accomodated at these sites. Also, catalytic mutants enabled the trapping of substrate in the +1 subsite, showing how Lam16A can hydrolyze both beta-1,4- and beta-1,3-glycosidic bonds.

Authors/Creators:Vasur, Jonas
Title:Probing promiscuity
Subtitle:structural studies of Phanerochaete chrysosporium Laminarinase 16A
Year of publishing :2009
Volume:2009:98
Number of Pages:36
Papers/manuscripts:
NumberReferences
ALLI Jonas Vasur, Rie Kawai, Anna M. Larsson, Kiyohiko Igarashi, Mats Sandgren, Masahiro Samejima and Jerry Ståhlberg (2006). X-ray crystallographic native sulfur SAD structure determination of laminarinase Lam16A from Phanerochaete chrysosporium. Acta Cryst D 62(Pt. 11), 1422–1429. II Jonas Vasur, Rie Kawai, Evalena Andersson, Kiyohiko Igarashi, Mats Sandgren, Masahiro Samejima and Jerry Ståhlberg (2009). X-ray crystal structures of Phanerochaete chrysosporium Laminarinase 16A in complex with products from lichenin and laminarin hydrolysis. FEBS Journal 276(14), 3858–3869. III Jonas Vasur, Rie Kawai, K. Hanna M. Jonsson, Göran Widmalm, Åke Engström, Martin Frank, Evalena Andersson, Henrik Hansson, Zarah Forsberg, Kiyohiko Igarashi, Masahiro Samejima, Mats Sandgren, Jerry Ståhlberg (2009). Synthesis of cyclic -glucan using Laminarinase 16A glycosynthase mutant from the basidiomycete Phanerochaete chrysosporium. (submitted to JACS in October) IV Jonas Vasur , Rie Kawai , Evalena Andersson , Mats Sandgren, Kiyohiko Igarashi , Masahiro Samejima , Jerry Ståhlberg (2009). Mimics of bound -1,3/1,6-glucan and lichenan in mutant Phanerochaete chrysosporium Laminarinase 16A. manuscript
Place of Publication:Uppsala
ISBN for printed version:978-91-576-7445-6
ISSN:1652-6880
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agrovoc terms:phanerochaete chrysosporium, hydrolases, beta glucanase, glucosides, enzyme activity, ligands, hydrolysis, analytical methods
Keywords:1, 3(4)-beta-D-glucanase, promiscuous, enzyme, protein-ligand structure, lichenan, lichenin, laminarin, glycoside hydrolase family 16
URN:NBN:urn:nbn:se:slu:epsilon-3026
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-3026
ID Code:2186
Department:(NL, NJ) > Dept. of Molecular Biology (until 131231)
Deposited By: Jonas Vasur
Deposited On:25 Nov 2009 00:00
Metadata Last Modified:02 Dec 2014 10:16

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