2018
DOI: 10.1074/jbc.ra117.000936
|View full text |Cite
|
Sign up to set email alerts
|

Identification of Euglena gracilis β-1,3-glucan phosphorylase and establishment of a new glycoside hydrolase (GH) family GH149

Abstract: Glycoside phosphorylases (EC 2.4.x.x) carry out the reversible phosphorolysis of glucan polymers, producing the corresponding sugar 1-phosphate and a shortened glycan chain. β-1,3-Glucan phosphorylase activities have been reported in the photosynthetic euglenozoan Euglena gracilis, but the cognate protein sequences have not been identified to date. Continuing our efforts to understand the glycobiology of E. gracilis, we identified a candidate phosphorylase sequence, designated EgP1, by proteomic analysis of an… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
3
2

Citation Types

5
47
1

Year Published

2018
2018
2024
2024

Publication Types

Select...
4
1

Relationship

3
2

Authors

Journals

citations
Cited by 34 publications
(53 citation statements)
references
References 59 publications
5
47
1
Order By: Relevance
“…The conserved structural characteristics include (a) the presence of a tryptophan ‐ asparagine ‐ aspartate (WND) motif (W651, N652, and D653) in the catalytic loop, with D653 as the predicted catalytic residue, although the D653 side chain is rotated 100° around CαCβ bond in comparison to that in PsLBP structure; (b) a conserved arginine ‐ aspartate (RD) motif (R470 and D471), which is involved in the recognition of sugar 1‐phosphate, although in the PsLBP structure, the distance between the aspartate residue (D375) and Glc1P is greater than the hydrogen bonding distance; (c) the conserved histidine (H959), which was predicted to be involved in phosphate recognition, although this residue does not form hydrogen bonds with the sulfate molecule in Pro_7066 or PsLBP structures. These structurally conserved amino acids are in agreement with the sequence alignment which predicted the presence of these residues in the enzyme active site . Bicine (BCN) derived from the crystallization precipitant was bound via hydrogen bonding contacts with R470 and E951, possibly mimicking the interaction of the enzyme active site with the hydroxy groups on C3 and C6 of Glc1P donor (Figure F).…”
Section: Resultssupporting
confidence: 77%
See 2 more Smart Citations
“…The conserved structural characteristics include (a) the presence of a tryptophan ‐ asparagine ‐ aspartate (WND) motif (W651, N652, and D653) in the catalytic loop, with D653 as the predicted catalytic residue, although the D653 side chain is rotated 100° around CαCβ bond in comparison to that in PsLBP structure; (b) a conserved arginine ‐ aspartate (RD) motif (R470 and D471), which is involved in the recognition of sugar 1‐phosphate, although in the PsLBP structure, the distance between the aspartate residue (D375) and Glc1P is greater than the hydrogen bonding distance; (c) the conserved histidine (H959), which was predicted to be involved in phosphate recognition, although this residue does not form hydrogen bonds with the sulfate molecule in Pro_7066 or PsLBP structures. These structurally conserved amino acids are in agreement with the sequence alignment which predicted the presence of these residues in the enzyme active site . Bicine (BCN) derived from the crystallization precipitant was bound via hydrogen bonding contacts with R470 and E951, possibly mimicking the interaction of the enzyme active site with the hydroxy groups on C3 and C6 of Glc1P donor (Figure F).…”
Section: Resultssupporting
confidence: 77%
“…The two copies of the molecule in the ASU form a biologically relevant homodimer with an interfacial area of ∼4230 Å 2 were calculated by jsPISA . The formation of a homodimer in the crystal structure is consistent with the gel filtration analysis, where Pro_7066 was eluted as a dimer . The domains present within each subunit can be defined as follows: an N‐terminal β‐sandwich (residues 1‐303, yellow), a helical linker region (residues 304‐341, lilac), an (α/α) 6 catalytic domain (residues 342‐1045, green) and a C‐terminal jelly roll domain (residues 1046‐1156, red) (Figure B).…”
Section: Resultssupporting
confidence: 67%
See 1 more Smart Citation
“…Inorganic phosphate, liberated as a result of glycosylation with Gal1P, can participate in the degradative phosphorolysis of β‐linked glucose disaccharide acceptors leading to in situ production of Glc1P (Figure ii). Given the greater kinetic efficiency with which natural donor Glc1P can be used by CDP and Pro_7066 (Table ), a series of extended glucose‐based oligosaccharides, together with glucose, might be formed in these reactions (Figure iii). Some of these oligosaccharides could become capped with galactose, thus leading to a mixture of products (Figure iv).…”
Section: Resultsmentioning
confidence: 99%
“…Following on from our efforts to understand GP structure–function relationships and their application in carbohydrate syntheses, herein we investigated the GH94 laminaribiose phosphorylase from Paenibacillus sp. YM‐1 ( Ps LBP), which has previously been reported for its specificity towards LB (β‐ d ‐glucopyranosyl‐(1→3)‐ d ‐glucopyranose; Scheme A) .…”
Section: Introductionmentioning
confidence: 99%