Folie 1

1
Multi-substrate flavonol O-glucosyltransferases
from strawberry (Fragaria x ananassa) achene and
receptacle Markus Griesser, Florian Vitzthum,
Mari Luz Bellido, Barbara Fink, Constanze Raasch,
Juan Munoz-Blanco, and Wilfried
Schwab Supplementary Data
A
B
8
8
6
6
4
4
2
2
rel. expression
rel. expression
0
0
-2
-2
-4
-4
-6
-6
4 h
6 h
8 h
4 h
6 h
8 h
C
D
8
9
7
8
7
6
6
5
5
4
rel. expression
rel. expression
4
3
3
2
2
1
1
0
0
4 h
6 h
8 h
4 h
6 h
8 h
Figure S1. Gene expression of FaGT6 (A, C) and
FaGT7 (B, D) in strawberry cell culture (A, B)
and strawberry fruit (C, D) after treatment with
salicylic acid () and in controls (?).
2
Supplemental data
D
60
50
40
mg MUG-equ./kg
30
20
10
0
2-naphthol
kaempferol
2-naphthol and kaempferol
Figure S2. Glucosylation of xenobiotics in
planta. Ripe strawberry fruits were injected with
10 mM 3-hydroxyflavone (A), 3-hydroxycoumarin (B)
and 2-naphthol (C). The fruits were harvested
after one day, extracted with methanol and
analyzed by LC-UV-ESI-MSn. The formation of the
respective glucosides were confirmed by product
ion experiments (MS2). Quantification of
glucosides derived from 2-naphthol and kaempferol
after injection of the individual alcohols and a
combination of the alcohols (10 mM each) (D).
Values are expressed as mg-equ./kg 4-methyl
umbelliferyl glucuronide (MUG). Levels of the
already formed kaempferol glucosides (23 mg
MUG-equ./kg) determined in untreated fruit were
substracted from the values obtained by the
treated fruits. The xenobiotic and the natural
substrate compete for the glucosyltransferase.
3
Figure S3. Mono view of conserved amino acids in
FaGT6, FaGT7, VvGT1, and MtGT71G1 shown in the
Michaelis complex of the Vitis vinifera
UDP-glucoseflavonoid 3-O-glycosyltransferase
with UDP-2-deoxy-2-fluoro glucose (green) and
kaempferol (red). Hydrogen bonds are indicated as
brown dotted lines. Amino acids close to
kaempferol are labeled (catalytically active
His23 and Asp128 as well as Phe130 and Leu156).
Figure was drawn with DeepView (Guex and Peitsch,
1997).
4
Met159
Met159
Ser159
His159
Figure S4. Comparison of amino acids located
close to the acceptor molecule in the active site
of four functionally characterized plant
glycosyltransferases which accept quercetin and
kaempferol. Amino acids in the vicinity (5 Å) of
kaempferol (pink) were identified in the
structure of VvGT and replaced by amino acids
that aligned to the identical positions in FaGT6,
FaGT7, and MtUGT71G1. UDP-glucose (green), amino
acids nonpolar (grey), basic (blue), acidic
(red), and polar (yellow). Figures were drawn
with DeepView (Guex and Peitsch, 1997).