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231223s2007 xx |||||o 00| ||eng c |
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|a 10.1111/j.1469-8137.2007.02174.x
|2 doi
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|a pubmed24n1253.xml
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|a (DE-627)NLM173932673
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|a (NLM)17888113
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Le Roy, Katrien
|e verfasserin
|4 aut
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| 245 |
1 |
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|a N-glycosylation affects substrate specificity of chicory fructan 1-exohydrolase
|b evidence for the presence of an inulin binding cleft
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| 264 |
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1 |
|c 2007
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| 336 |
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|a Text
|b txt
|2 rdacontent
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| 337 |
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|a ƒaComputermedien
|b c
|2 rdamedia
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| 338 |
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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| 500 |
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|a Date Completed 13.12.2007
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|a Date Revised 09.01.2024
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|a published: Print
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|a Citation Status MEDLINE
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|a Recently, the three-dimensional structure of chicory (Cichorium intybus) fructan 1-exohydrolase (1-FEH IIa) in complex with its preferential substrate, 1-kestose, was determined. Unfortunately, no such data could be generated with high degree of polymerization (DP) inulin, despite several soaking and cocrystallization attempts. Here, site-directed mutagenesis data are presented, supporting the presence of an inulin-binding cleft between the N- and C-terminal domains of 1-FEH IIa. In general, enzymes that are unable to degrade high DP inulins contain an N-glycosylation site probably blocking the cleft. By contrast, inulin-degrading enzymes have an open cleft configuration. An 1-FEH IIa P294N mutant, introducing an N-glycosylation site near the cleft, showed highly decreased activity against higher DP inulin. The introduction of a glycosyl chain most probably blocks the cleft and prevents inulin binding and degradation. Besides cell wall invertases, fructan 6-exohydrolases (6-FEHs) also contain a glycosyl chain most probably blocking the cleft. Removal of this glycosyl chain by site-directed mutagenesis in Arabidopsis thaliana cell wall invertase 1 and Beta vulgaris 6-FEH resulted in a strong decrease of enzymatic activities of the mutant proteins. By analogy, glycosylation of 1-FEH IIa affected overall enzyme activity. These data strongly suggest that the presence or absence of a glycosyl chain in the cleft is important for the enzyme's stability and optimal conformation
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a Arabidopsis Proteins
|2 NLM
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| 650 |
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|a Plant Proteins
|2 NLM
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| 650 |
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|a Inulin
|2 NLM
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|a 9005-80-5
|2 NLM
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| 650 |
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|a Glycoside Hydrolases
|2 NLM
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| 650 |
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|a EC 3.2.1.-
|2 NLM
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| 650 |
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|a Inv1 protein, Arabidopsis
|2 NLM
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| 650 |
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|a EC 3.2.1.26
|2 NLM
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| 650 |
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|a beta-Fructofuranosidase
|2 NLM
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| 650 |
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|a EC 3.2.1.26
|2 NLM
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| 650 |
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|a fructan beta-fructosidase
|2 NLM
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| 650 |
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|a EC 3.2.1.80
|2 NLM
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| 700 |
1 |
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|a Verhaest, Maureen
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Rabijns, Anja
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Clerens, Stefan
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Van Laere, André
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Van den Ende, Wim
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t The New phytologist
|d 1979
|g 176(2007), 2 vom: 17., Seite 317-324
|w (DE-627)NLM09818248X
|x 1469-8137
|7 nnns
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| 773 |
1 |
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|g volume:176
|g year:2007
|g number:2
|g day:17
|g pages:317-324
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| 856 |
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|u http://dx.doi.org/10.1111/j.1469-8137.2007.02174.x
|3 Volltext
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|a AR
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|d 176
|j 2007
|e 2
|b 17
|h 317-324
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