Supplementary Figures and Table
Fig. S1. Identification of the dimeric TGase 2 region.(A) TGase 2 was incubated without or with CaCl2 during 30 min at 20 or 37 °C. The reactants were resolved by 10% native gel and stained with coomassieblue. (B) LC-MS/MS analysis of trypsin-digested monomer and dimer of TGase 2 from (A) gel. The mass coverage from sample shown the distinction between monomer and dimer of TGase 2. Confirmed sequences are marked in green (high confidence) and yellow (medium confidence), while unidentified regions are marked in red. The blue under-line denotes the region (593ILGEPKQK600) facing interdimericTGase 2.
Fig. S2. Schematic representation of the dimeric TGase 2 regions facing each other.(A) Crystal structure of the open conformation of TGase 2 (PDB: 2Q3Z). TGase 2 is composed of four domains, namely, the N-terminal domain (azure), catalytic domain (verdigris), C-terminal domain 1 (green), and C-terminal domain 2 (yellow). The redbelt within the C-terminal domain 2 denotes the region (593ILGEPKQK600) facinginterdimericTGase 2 by MS analysis. (B) Sequence chain view of TGase 2 with the dimer interfacing region by RCSB PDB ( The purple, pink, and red curves represent turns, 3/10-helices, and -helices, respectively. The yellow and gold arrowheads represent -strands and -bridges, respectively. The blue lines represent bends, and the black lines represent no specific secondary structure. The red dotted-line rectangle denotes the region (593ILGEPKQK600) facinginterdimericTGase 2. SCOP represent a database of structural classification of proteins ( DSSP is a database of secondary structure assignment for all protein entries ( in the Protein Data Bank (PDB).
Table S1. Summary of TGase 2 conformation related function.
Singh et al[25] / Kim et al(this study)TGase 2 type / With/without GTP / With calcium / With/without
GTP / With calcium
Wild type TGase 2 / With GTP
Folded form
Catalytic inactive / Unfolded form
Catalytic active / With GTP
Folded from
(data not shown)
Catalytic inactive / Unfolded form
Catalytic active in 2 types.
- Intra crosslink under 30 C,
- Inter crosslink over 30 C
Without GTP
ND / Without GTP
Unfolded form
In 2 types.
- Monomer form under 30 C
- Dimer form over 30 C
Catalytic inactive
Mutant TGase 2 (R580K) / (similar to without GTP condition - GTP binding loss)
unfolded form
activity ND / Unfolded form
Catalytic active / ND / ND
Deletion mutant TGase 2 (1-548) / unfolded form
Catalytic inactive / unfolded form
Catalytic inactive / ND / ND
ND, not determined