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2025-09-16
Cross-linking mass spectrometry experimental design
LC-IMS-MS/MS sample description

Cross-linking mass spectrometry experimental design


Scheme of photoaffinity labeling. NPY containing photo-leucines at positions 17, 24, and 30 was crosslinked to bicelle-reconstituted Y2R preparations by UV-A light. The crosslinked complex is enzymatically digested, peptides are analyzed by LC-IMS-MS/MS using the MeroX software.




LC-IMS-MS/MS sample description


Replicate Sample ID Group Acquisition Order Condition Chromatography Condition MS Mode
timsTOF_090822_pMet_Y2R_no_pNPY S-Trap Control (low CE; 20 µg Y2R-bicelle) S-Trap Control (low CE; 20 µg Y2R-bicelle) 3389 Y2R-Bicelle Control µPAC (90 min) DDA-PASEF
timsTOF_090822_pMet_Y2R-pNPY_XL_1 S-Trap XL (low CE; 20 µg Y2R-bicelle) 1 S-Trap XL (low CE; 20 µg Y2R-bicelle) 3391 Y2R-Bicelle

µPAC (90 min)

DDA-PASEF
timsTOF_090822_pMet_Y2R-pNPY_XL_2 S-Trap XL (low CE; 20 µg Y2R-bicelle) 2 S-Trap XL (low CE; 20 µg Y2R-bicelle) 3393 Y2R-Bicelle µPAC (90 min) DDA-PASEF
timsTOF_090822_pMet_Y2R-pNPY_XL_3 S-Trap XL (low CE; 20 µg Y2R-bicelle) 3 S-Trap XL (low CE; 20 µg Y2R-bicelle) 3395 Y2R-Bicelle µPAC (90 min) DDA-PASEF
timsTOF_090922_pMet_Y2R-pNPY_XL_4 S-Trap XL (high CE; Y2R-bicelle) 1 S-Trap XL (high CE; Y2R-bicelle) 3521 Y2R-Bicelle µPAC (90 min) DDA-PASEF
timsTOF_2010922_N-term-Y2R_1-45-no_pNPY S-Trap Control (low CE; 20 µg N-term) 1 S-Trap Control (low CE; 20 µg N-term) 3628 NT-Y2R Control Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_2010922_N-term-Y2R_1-45-pNPY_XL_1 S-Trap XL (20 µg NT-Y2R) 1 S-Trap XL (20 µg NT-Y2R) 3631 NT-Y2R Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_2010922_N-term-Y2R_1-45-pNPY_XL_2 S-Trap XL (20 µg NT-Y2R) 2 S-Trap XL (20 µg NT-Y2R) 3632 NT-Y2R Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_2010922_N-term-Y2R_1-45-pNPY_XL_3 S-Trap XL (20 µg NT-Y2R) 3 S-Trap XL (20 µg NT-Y2R) 3633 NT-Y2R Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_20221026_N-term-Y2R_1-45-pNPY_XL_1_low-CE S-Trap XL (low CE; 20 µg NT-Y2R) 1 S-Trap XL (low CE; 20 µg NT-Y2R) 3655 NT-Y2R Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_20221026_N-term-Y2R_1-45-pNPY_XL_2_low-CE S-Trap XL (low CE; 20 µg NT-Y2R) 2 S-Trap XL (low CE; 20 µg NT-Y2R) 3656 NT-Y2R Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_20221026_N-term-Y2R_1-45-pNPY_XL_3_low-CE S-Trap XL (low CE; 20 µg NT-Y2R) 3 S-Trap XL (low CE; 20 µg NT-Y2R) 3657 NT-Y2R Self-Packed (3-50; 90 min) DDA-PASEF
timsTOF_20230129_N-term-Y2R_1-45-pNPY_XL_DIA_1 S-Trap XL (DIA-PASEF; 20 µg NT-Y2R) 1 S-Trap XL (DIA-PASEF; 20 µg NT-Y2R) 3920 NT-Y2R Self-Packed (3-50; 90 min) DIA-PASEF
timsTOF_20230129_Y2R-pNPY_DIA_01 S-Trap XL (DIA-PASEF; 20 µg Y2R-bicelle) 1 S-Trap XL (DIA-PASEF; 20 µg Y2R-bicelle) 3921 Y2R-Bicelle Self-Packed (3-50; 90 min) DIA-PASEF
timsTOF_20230302_Y2R-pNPY_XL S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 4101 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20230302_N-term-Y2R_1-45-pNPY_XL S-Trap XL (low CE; 20 µg NT-Y2R) long gradient 1 S-Trap XL (low CE; 20 µg NT-Y2R) long gradient 4102 NT-Y2R Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_JNJ_1 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8683 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_JNJ_2 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8684 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_JNJ_3 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8685 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_NPY_1 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8687 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_NPY_2 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8688 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_NPY_3 S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8689 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF
timsTOF_20241215_Y2R-pNPY_no_UV S-Trap XL (low CE; 20 µg Y2R-bicelle) long gradient 1   8691 Y2R-Bicelle Self-Packed (3-35; 90 min) DDA-PASEF