Covalent binding affinity to VAT1 at Cys86 residue in human NCI-H358 cells assessed as log2 H/L ratio at 10 uM pretreated for 3 hrs followed by N-(((6S,7R)-7-(4-chloro-3-fluorophenyl)-1,4-oxazepan-6-yl)methyl)-2-hydroxyacetamide treatment for 3 hrs by competitive TMT click chemistry pull-down experiment based LC-MS/MS analysis
Covalent binding affinity to VAT1 at Cys residue in human NCI-H358 cells assessed as reduction of log2 H/L ratio for tryptic peptide ACGLNFADLMAR incubated for 4 hrs followed by cell lysis and subsequently labelled with light thiol probe N-((S)-18-iodo-11-isopropyl-10,13,17-trioxo-3,6-dioxa-9,12,16-triazaoctadecyl)-6-((4R,5S)-5-methyl-2-oxoimidazolidin-4-yl)hexanamide and heavy thiol probe 4 for 1 hr under dark conditions followed by tryptic digestion for overnight and measured by discovery ABPP based nano LC-MS/MS analysis
Covalent binding affinity to VAT1 at Cys residue in human NCI-H358 cells assessed as reduction of log2 H/L ratio for tryptic peptide CLVLTGFGGYDK incubated for 4 hrs followed by cell lysis and subsequently labelled with light thiol probe N-((S)-18-iodo-11-isopropyl-10,13,17-trioxo-3,6-dioxa-9,12,16-triazaoctadecyl)-6-((4R,5S)-5-methyl-2-oxoimidazolidin-4-yl)hexanamide and heavy thiol probe 4 for 1 hr under dark conditions followed by tryptic digestion for overnight and measured by discovery ABPP based nano LC-MS/MS analysis