Journal: JIMD Reports

Article Title: Small Molecules as Alternate Substrates for 3‐Methylglutaconylation

doi: 10.1002/jmd2.70047

Figure Lengend Snippet: Non‐enzymatic reactions of trans‐3MGC‐CoA. The series of non‐enzymatic chemical reactions induced by the accumulation of trans ‐3MGC‐CoA in vivo is depicted. Trans ‐3MGC‐CoA formed as an intermediate in the leucine catabolism pathway or alternatively by the acetyl CoA diversion pathway spontaneously isomerizes to cis ‐3MGC‐CoA. A subsequent intramolecular cyclization reaction generates cis ‐3MGC anhydride and free CoA. 3MGC anhydride has three potential fates, including hydrolysis to yield cis ‐3MGC acid (center), acylation of protein lysine residues (left), or acylation of amino group‐containing small molecules (right).

Article Snippet: 3‐MGC anhydride (4‐methyl‐2H‐pyran‐2,6(3H)‐dione) was from Biosynth Ltd. (UK).

Techniques: In Vivo

Journal: JIMD Reports

Article Title: Small Molecules as Alternate Substrates for 3‐Methylglutaconylation

doi: 10.1002/jmd2.70047

Figure Lengend Snippet: Effect of glycine on the signal intensity of 3MGCylated BSA. (A) Schematic diagram depicting the experimental design employed. “Incubation 1” refers to the incubation of 3MGC‐CoA filtrate with glycine while “Incubation 2” refers to the further incubation of this sample following the addition of BSA. Dashed box represents expected products formed. Control experiments with no glycine added were conducted in parallel. (B) Incubations were conducted as described in panel A and, following incubation, an aliquot of each sample was subjected to SDS‐PAGE and immunoblotting with α‐3MGC IgG. Dotted line indicates that the immunoblot was cut to remove extraneous lanes. (C) Incubations were conducted and processed as in panel B, except 3MGC anhydride was used in lieu of trans ‐3MGC‐CoA filtrate.

Article Snippet: 3‐MGC anhydride (4‐methyl‐2H‐pyran‐2,6(3H)‐dione) was from Biosynth Ltd. (UK).

Techniques: Incubation, Control, SDS Page, Western Blot

Journal: JIMD Reports

Article Title: Small Molecules as Alternate Substrates for 3‐Methylglutaconylation

doi: 10.1002/jmd2.70047

Figure Lengend Snippet: Effect of small molecules on trans ‐3MGC‐CoA‐dependent acylation of BSA. (A) A trans ‐3MGC‐CoA‐containing 3‐MCC reaction filtrate was incubated with indicated amounts of N‐acetylglucosamine or glucosamine for 2 h at 37°C. BSA (0.5 mg/mL) was then added, and the samples incubated for a further 24 h. An aliquot of each sample was then subjected to α‐3MGC IgG immunoblot analysis. (B) A trans ‐3MGC‐CoA‐containing 3‐MCC reaction filtrate was incubated with specified amounts of choline or ethanolamine for 2 h at 37°C. BSA (0.5 mg/mL) was then added, and the samples incubated for a further 24 h. An aliquot of each sample was then subjected to SDS‐PAGE and immunoblotting with α‐3MGC IgG. (C) A trans ‐3MGC‐CoA‐containing 3‐MCC reaction filtrate was incubated with specified amounts of glutathione for 2 h at 37°C. BSA (0.5 mg/mL) was then added, and the samples incubated for a further 24 h. An aliquot of each sample was then subjected to α‐3MGC IgG immunoblot analysis.

Article Snippet: 3‐MGC anhydride (4‐methyl‐2H‐pyran‐2,6(3H)‐dione) was from Biosynth Ltd. (UK).

Techniques: Incubation, Western Blot, SDS Page