Figure 2: Y26 phosphorylation promotes cofactor 2,3-BPG binding to PGAM1. | Nature Communications

Figure 2: Y26 phosphorylation promotes cofactor 2,3-BPG binding to PGAM1.

From: Tyr26 phosphorylation of PGAM1 provides a metabolic advantage to tumours by stabilizing the active conformation

Figure 2

(a) Cartoon representation of 2,3-BPG location from structure 3FDZ superposed on PGAM1 (PDB accession code: 1YFK). H11 and Y92 are directly proximal to and Y26 is also close to cofactor (2,3-BPG)/substrate (3-PG) binding site. (b) 2,3-BPG analogue (8-hydroxy-1,3,6-pyrenetrisulfonate) competes with 2,3-BPG for binding to rPGAM1 protein, where 3 μM 2,3-BPG analogue was mixed with different concentrations of 2,3-BPG in reaction mixture containing 100 mM Tris-HCl (pH 7.5). Fluorescence intensity of 2,3-BPG analogue (ex: 362 nm, em: 520 nm) was measured before and 5 min after the addition of 2.3 μM rPGAM1 protein to the reaction mixture. Decrease in fluorescence intensity of 2,3-BPG analogue indicates 2,3-BPG analogue binding to rPGAM1 protein. The values are presented as relative fluorescence units (RFU). (c) Purified Flag-PGAM1 WT or Y26F were incubated with cofactor (represented by 2,3-BPG analogue, 8-hydroxy-1,3,6-pyrenetrisulfonate). The cofactor binding affinity was determined by decrease in fluorescence intensity of the analogue. The values are presented as RFU. The error bars represent mean values±s.d. from three independent experiments (*0.01<P<0.05; **P<0.01).

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