Cell News 02/2017
12
NIKON YOUNG SCIENTIST AWARD 2017
Uninfected
Lys0 Arg0
S
.
Typhimurium
Lys8 Arg10
K-GG
K-GG
GG-K
anti-K-GG IP
LC-MS
Data analysis
m/z
Intensity
LysC / Trypsin digest
K-GG
K-GG
GG-K
K
K
K48
M1
Trypsin digest
anti-K48 pUb IP
K-GG
K-GG
GG-K
anti-M1 pUb IP
Trypsin digest
4.5
5.5
6.5
7.5
8.5
pH
SCX
LC-MS
Data analysis
m/z
Intensity
LC-MS
Data analysis
m/z
Intensity
K-GG
K-GG
GG-K
# of regulated ubiquitination sites
Log 2 (H:L)
-2.5
5
4
5
1
3 2
≥6
UBE2L3
TNIP1
RELA
TRIM25
IKBKG
BIRC2
NFKB1
TRAF2
SHARPIN
FAS
RBCK1
FAM105B
TRAF6
RNF31
PRKCI
TOM1
STAM
ANXA1
SH3BP4
STAM2 TFRC
RAB14
COPA
STOM
RALA
EPS15 RAB10
SNX9
HSPA8
NEDD4L
PDCD6IP
HSPA6
CLTC
EPN1
EGFR
ATP6V1H
ATP6V0A1
CLINT1
ARHGAP29
CDC42
GRTP1
RHOG
PI4KA
PIP5K1A
PIP5K1C
DAG1
ARHGAP1
RAC1
ABI2
MSN
BCAR1
ITGB1
EZR
RDX
PPP1R1A2
ITGB4
STAT3
CAPN1
BNIP2
IQGAP1
PFN1
ARHGDIA
MYO1B
NRAS
MYO1E
IQGAP2
SH3RF1
WASL
Regulation of apoptosis
Regulation of actin cytoskeleton
Internal side of plasma membrane
a
b
Figure 1: Global Analysis of the Ubiquitinome in human epithelial cells infected with Salmonella typhimurium
(a) Experimental workflows of complementary SILAC-based ubiquitin proteomics methods used to monitor changes in the ubiquitination status of proteins upon
Salmonel-
la
infection. (b) Known protein-protein interactions of protein hits harboring regulated ubiquitination sites (log2 (H:L) ratio >1 or <-1). Hits were clustered based on proces-
ses found over-represented in gene ontology enrichment analysis. Node size and color for each protein indicate the number of regulated diGly sites and their mean log2
(H:L) ratio, respectively. Adapted from Fiskin
et al
., 2016
