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Lei C, Chen Z, Hao Y, Huang W, Chu T, Xiao K, Zhang C, Zhou W, Li C, Chen X. Quantitative and site-specific chemoproteomic profiling of O-GlcNAcylation in Drosophila. Bioorganic & medicinal chemistry 2025 124 40245499
Abstract:
Protein O-GlcNAcylation plays a crucial role in Drosophila melanogaster development. Dysregulation of O-GlcNAc transferase (sxc/Ogt) and O-GlcNAcase (Oga) disrupts early embryogenesis and locomotor behavior. It is therefore of great interest to identify and quantitatively analyze O-GlcNAcylation sites in Drosophila. Here, we perform quantitative and site-specific profiling of O-GlcNAcylation in Drosophila by employing a chemoenzymatic labeling strategy. A total of 2196 unambiguous O-GlcNAcylation sites and 1308 O-GlcNAcylated proteins are identified. Quantitative analysis of O-GlcNAcylation in the head of Drosophila with sxc/Ogt knockdown in GABAergic neurons reveals a reduction in O-GlcNAcylation of several proteins involved in muscle development, consistent with the phenotypic defects observed in sxc/Ogt RNAi Drosophila. Furthermore, quantitative analysis of O-GlcNAcylation under a high-sugar diet reveals altered O-GlcNAcylation of several proteins associated with obesity and neurological diseases, such as Hex-A and Ankyrin 2. Our study not only establishes an effective method for large-scale identification of O-GlcNAcylation sites, but also provides a valuable resource for studying O-GlcNAc biology in Drosophila.
O-GlcNAc proteins:
A0A075BK61, A0A075BLW4, A0A0B4JCT8, A0A0B4JD24, NEP2, A0A0B4K6D6, A0A0B4K6H8, A0A0B4K6N6, A0A0B4K707, A0A0B4K7F6, RBP2, A0A0B4K7K9, A0A0B4K7Y7, A0A0B4K851, A0A0B4KEI6, A0A0B4KEK1, A0A0B4KEN1, A0A0B4KEP4, A0A0B4KEQ0, A0A0B4KEQ1, A0A0B4KER6, A0A0B4KEW3, A0A0B4KEY5, A0A0B4KF23, A0A0B4KF57, A0A0B4KFC5, A0A0B4KFE3, A0A0B4KFF4, A0A0B4KFI9, A0A0B4KFJ8, A0A0B4KFK9, A0A0B4KFP4, A0A0B4KFY9, A0A0B4KFZ2, A0A0B4KG52, A0A0B4KGP0, A0A0B4KGP2, A0A0B4KGX7, A0A0B4KH25, A0A0B4KH51, A0A0B4KHB2, A0A0B4KHE7, A0A0B4KHJ7, A0A0B4KHX4, A0A0B4KHY8, A0A0B4KI07, A0A0B4KI32, A0A0B4KI37, A0A0B4KI71, A0A0B4LEW1, A0A0B4LEY9, A0A0B4LF06, A0A0B4LF60, A0A0B4LF93, A0A0B4LFB3, A0A0B4LFH2, A0A0B4LFP7, A0A0B4LFX4, A0A0B4LG05, A0A0B4LG23, A0A0B4LG93, A0A0B4LGJ1, A0A0B4LGS5, A0A0B4LH64, A0A0B4LHK4, A0A0B4LHS9, A0A0B4LHV0, A0A0B4LIF3, A0A0B4LIM4, A0A0B4LJ24, A0A0C4DHC2, A0A0H4Y1G0, A0A0P0I6H8, A0A0S0WNG8, A0A0S0X7W8, A0A126GUS1, A0A126GUT4, A0A165Q6N5, A0A1B2AJZ7, A1A6T6, A1Z7S3, A1Z8M2, A1Z968, A1Z9J3, A1Z9M5, A1ZAD3, A1ZAD5, A1ZAG9, A2VEH2, A2VEJ3, A2VEN0, A2VES6, A4IJ75, A4V0A1, A4V166, A4V193, A4V1B2, A4V1N8, A4V1N9, A4V201, A4V379, A4V3L3, A4V3N7, A4V449, A5JLB7, A7LBP7, A8DY82, A8DY98, A8DYD4, OBSCN, A8DYS7, A8DZ25, A8DZ29, A8E6M3, A8JNT5, A8JNU1, A8JQU7, A8JUU3, SBNO, A8JV07, A8JV18, A8WHE9, A8WHF5, A8WHG6, A8WHH5, A8WHL8, A8Y4V5, A9UNF4, A9YGT1, A9YHJ2, A9YIJ2, B3DMQ0, B3DN78, B3DNL9, B3LVC7, B4F582, B5RIV2, B5RIV5, B5RJ67, B5RJR0, B5X535, B7FNR3, B7YZL8, B7YZQ7, B7YZR7, B7YZS3, B7Z086, B7Z090, B7Z0D4, B7Z0H1, B7Z0K4, B8A408, B8A429, B9EQY8, Y5076, FAU, C0MII8, C0MLZ0, C0PTX6, C1C5B5, C3KGJ2, C3KGM0, C3KKB9, C4IXY4, C4IXY9, C4IXZ0, C6SUY0, C6SV07, C6TP30, C7LA61, C7LA69, C7LA93, C7LAB5, C7LAC4, C9QP08, C9QPG6, C9QPJ3, D0IQG1, D0IQJ5, D0Z730, D0Z760, D1Z3A5, D2NUL5, D3DMC9, D3DMH7, D3DN25, D5A7R4, D5AEJ6, D5SHS6, D5SHV0, D7FA76, D9IWU3, E0R7M1, E0R908, E0R956, E1JGK6, E1JGP4, E1JGV4, E1JH30, E1JH57, E1JHV7, E1JI01, E1JIG1, E1JIJ9, E1JIT4, E1JIZ1, E1JJ59, E1NZB6, E1NZD0, E1NZD7, E2QC56, E2QCL8, E2QCY9, E2QCZ3, E2QCZ8, E2QD61, E2QD65, E2QD73, E3CTP7, E3CTQ4, E4NKH3, E4NKJ7, E6EK15, E8NH10, E8NH12, E8NH67, E8NH84, E9P245, F0JAJ0, F0JAT3, F2FBA6, F3YD57, F3YDJ9, F3YDK4, F6J363, F6J412, F6J5C6, F6J5L2, F6J6M1, F9W345, G2J5V2, G3JX33, G4LU05, G4LU55, H0RNE3, H1UUC4, H1UUG6, H1UUH1, H1ZY75, H1ZYH2, H5V8A7, H5V8C9, H5V8D4, H6QXJ8, H9XVM0, H9XVM9, H9XVP3, I1V4X8, I6QP77, L0CRN5, L0MLM5, L0MPW8, M9MRD4, M9MRJ4, M9MRR7, M9MRX4, M9MS06, M9MSF4, M9MSF5, M9MSG3, M9NCS8, M9NCX1, M9NDC9, M9NDM3, M9NDT7, M9NE59, M9NE82, M9NED8, M9NEL3, M9NFP9, M9NFY9, M9NGG5, M9NGK3, M9NGY0, M9PB99, M9PBA9, M9PBG8, M9PBH6, M9PBL6, M9PBM2, M9PBM5, M9PBU4, M9PBV6, M9PBZ3, M9PBZ9, M9PC89, M9PCE8, M9PCI5, M9PCJ8, M9PCK1, M9PCQ6, M9PCS5, M9PD58, M9PDC7, M9PDE2, M9PDL8, M9PDW3, M9PDX8, M9PE03, M9PE50, M9PE74, M9PE86, M9PEA9, M9PEB1, M9PEF6, M9PEG8, M9PEI7, M9PEK0, M9PEK8, M9PEP8, M9PEZ0, M9PF16, M9PF54, M9PF71, M9PF74, M9PFB2, M9PFE5, M9PFM9, M9PFP3, M9PFU6, M9PFW3, M9PG32, M9PG55, M9PGA2, M9PGG4, M9PGT0, M9PHG4, M9PHL0, M9PHZ2, M9PI47, M9PIB5, M9PIF1, M9PJ79, M9PJC7, N0D6X7, O16804, O18354, O18363, ITA3, O46067, O61348, RAB7, O76904, O77284, O77434, O96607, ZEN1, RL4, ROUGH, LAMB1, ITA2, PCNA, TRX, RCC1, HMPB, PTP10, LOLA2, COPB, ACTZ, OB69A, HASP, SGS4, HMSH, Q058R9, Q058U1, Q058W4, Q06AJ1, Q06PM6, Q0E8S7, Q0E8T4, Q0E9E6, Q0E9I8, Q0GA04, Q0KHZ6, DMDD, Q1EBZ0, Q1EC05, Q1EC11, Q1NZ60, Q1RKW2, Q1RKX3, Q1WWA1, Q1WWC0, Q1WWE5, Q1WWG0, Q1WWG9, Q23980, Q24043, Q24107, TFDP, Q29QN4, Q29QZ2, Q29R35, Q29R42, Q2L385, Q2MGK5, Q2PNQ2, Q2XY62, Q3ZAJ5, Q45EX0, Q4QQ20, Q4V3J4, Q4V3Q9, Q4V3S6, Q4V3T3, Q4V3X8, Q4V438, Q4V451, Q4V4Y8, Q4V512, Q4V5A7, Q4V5E9, Q4V5T6, Q4V6F3, Q4V6N1, Q4V6U5, Q53YH4, Q58CJ5, Q58CK8, Q58S76, Q58SM2, Q59DZ3, Q59E01, Q5BI73, Q5BIJ2, Q5BIL5, ATM, Q5U0U9, Q5U0W8, Q5UHE2, Q6AWJ8, Q6AWS1, Q6GUS1, Q6IDG5, Q6IG67, Q6IHA6, Q6IIH5, Q6IIJ2, Q6IJK3, Q6IKL5, Q6IKR1, Q6IL25, Q6ILA9, Q6ILJ5, Q6NL43, Q6NL44, Q6NL84, Q6NLM6, Q6NN63, Q6NND6, Q6NNF8, Q6NP08, Q6NP20, Q6NP25, Q6NP27, Q6NP54, Q6NP56, Q6NP81, Q6NR17, BOCKS, Q711U4, Q7JP25, Q7JQK6, Q7JQL5, Q7JRF0, EAF, Q7JW07, NUP62, SEELE, Q7JYQ4, NUP50, Q7K0K1, Q7K105, Q7K126, Q7K180, Q7K1C5, Q7K1M7, Q7K2L3, Q7K2Q6, SEH1, Q7K3E2, Q7K3X8, Q7K3Z3, Q7K501, Q7K554, SCARF, Q7KHK9, Q7KLE5, Q7KMJ0, Q7KMN4, Q7KN04, GGYF1, Q7KS07, Q7KSB7, Q7KT11, Q7KTA1, Q7KTG5, Q7KTU3, Q7KU92, Q7KUB5, Q7KUK9, Q7KV35, Q7KV90, DCLK, INT3, Q7YU14, Q7YU46, Q7YU50, Q7YU82, Q7YU85, Q7YU88, LOLA4, PPN, Q86BA5, Q86BH5, Q86BH6, Q86BM5, Q86BS0, Q86LF3, Q86LF6, Q86MQ2, Q86NN4, Q86NV5, Q86P46, Q86P90, Q86PF3, Q86R99, Q8IGF3, Q8IGV8, Q8IH74, Q8IHB0, Q8IHF0, Q8IMB7, Q8IMB8, Q8IMB9, Q8IMH4, Q8IMJ3, Q8IMU1, Q8INA9, Q8INN5, DAAF1, Q8INU6, Q8INW9, Q8INX7, Q8IPP9, UBP2, Q8IQN2, Q8IQN7, Q8IQQ7, Q8IQT4, Q8IQT9, Q8IQW5, Q8IQX5, Q8IR52, Q8IR57, Q8IRC1, Q8IRQ6, TRR, LBR, Q8MLV5, Q8MLW2, Q8MQI1, Q8MQJ3, Q8MQN5, Q8MQW5, Q8MQY1, Q8MR22, Q8MR81, Q8MRE8, Q8MRI4, Q8MRL5, Q8MRM4, Q8MRN0, Q8MRS3, Q8MRY4, Q8MS79, Q8MSM1, Q8MSP6, Q8MSQ2, Q8MSS9, SYMPK, SHEP, Q8MT64, Q8MT92, FACD2, Q8SWT1, Q8SX39, Q8SX64, Q8SX82, Q8SX92, Q8SXA3, Q8SXE4, Q8SXF2, Q8SXR0, Q8SXU9, GAWKY, Q8SY39, Q8SYJ0, Q8SYT3, Q8SZK5, Q8SZW9, Q8T038, Q8T076, Q8T0C9, Q8T0H7, Q8T0N3, Q8T0U9, Q8T3H4, Q8T3I6, Q8T3Z0, Q8T419, Q8T435, Q8T484, Q8T4F8, Q8T4G0, Q8T8S6, Q8T8V6, Q8T8Z1, Q8T8Z9, Q8T921, Q8T9B4, Q8T9D1, Q8T9H1, Q8T9J2, Q94885, Q95R75, GILT1, Q95RK8, Q95RM4, Q95RS3, Q95RT8, KCMF1, Q95S27, Q95S58, Q95SF4, Q95SR0, Q95SX4, Q95T04, Q95T29, Q95TA4, Q95TC3, Q95TF7, Q95TJ6, Q95TP6, Q95TR0, Q95TS6, Q95TZ0, Q95TZ1, Q95TZ7, Q95U09, Q95U36, Q95U61, Q960C2, LRCH, Q960F4, Q960G3, Q960J1, Q960L2, Q960M4, Q960S0, Q960V1, FACR3, Q960X2, Q960Y7, Q961D0, Q961F0, Q961F9, Q961K6, Q961P7, Q961R5, Q961T0, Q99140, Q9BII5, Q9BPQ2, Q9BPQ3, Q9GPN8, Q9GQA3, Q9GT73, Q9I7L8, LAR4, Q9N9Z1, Q9N9Z2, Q9NG60, Q9NGV3, Q9NGZ4, SAN, CAPS, Q9NHF7, Q9NHX5, NXF1, Q9U4G5, Q9U4I2, Q9U5E2, Q9U6X3, Q9U916, Q9V3C8, Q9V3I0, SP27A, Q9V3U6, Q9V3Z3, Q9V430, Q9V445, HCF, MED26, LOLA5, Q9V9R2, Q9V9U9, Q9VAJ1, Q9VB52, Q9VBK6, Q9VBK9, Q9VC19, Q9VC36, Q9VC39, NUP98, Q9VCL9, Q9VCR9, DCR1, Q9VD85, FANCM, Q9VDD0, Q9VDF4, Q9VDI1, Q9VDN6, Q9VDP8, Q9VDU7, NUP58, Q9VE37, Q9VEW1, Q9VEZ2, Q9VF20, Q9VF39, Q9VF65, Q9VFA8, Q9VFC1, Q9VG26, Q9VGG9, Q9VGJ2, Q9VGL0, Q9VH03, Q9VH10, Q9VH23, Q9VH63, Q9VH64, Q9VH97, Q9VHC0, Q9VHC8, Q9VI26, Q9VI53, Q9VI61, Q9VIE0, Q9VIF2, Q9VIN9, SWM, Q9VJD1, Q9VJH6, Q9VJK1, Q9VJX4, Q9VJY7, Q9VK82, Q9VKC6, Q9VKU0, Q9VKZ7, Q9VL05, Q9VL29, Q9VL48, Q9VL91, Q9VLB4, Q9VLP2, Q9VMD2, DNAT2, Q9VMR6, Q9VMS1, Q9VMS2, ITPA, Q9VN08, Q9VNE0, Q9VNI5, Q9VNR6, Q9VNT0, Q9VNU3, Q9VP27, Q9VP43, Q9VP57, Q9VPA9, ATK, Q9VPF6, Q9VPG2, Q9VPR2, Q9VQ58, NPC2, Q9VQE6, Q9VQM0, Q9VQP5, Q9VQR3, Q9VQS4, Q9VQU8, Q9VQZ3, Q9VRE0, Q9VRM6, Q9VS01, Q9VS67, CP316, Q9VS82, Q9VSA5, Q9VSN2, Q9VSN5, ATAT1, JARD2, Q9VT61, Q9VTR6, KRI1, VP13D, DBNL, Q9VUB1, Q9VUB5, FRL, Q9VUG6, Q9VUH6, Q9VUH8, Q9VV52, MINP1, Q9VVF1, Q9VVU1, Q9VW05, Q9VW34, Q9VW82, Q9VWA6, Q9VWA7, Q9VWG1, Q9VWJ3, Q9VWN2, Q9VWP8, Q9VWY8, TM41, Q9VXA3, Q9VXF4, Q9VY22, Q9VY46, Q9VYH8, Q9VYJ1, Q9VYK0, Q9VZ30, Q9VZ67, Q9VZE8, ACK, Q9VZX8, Q9W002, Q9W061, Q9W0U4, Q9W124, Q9W125, Q9W140, Q9W158, Q9W170, Q9W194, Q9W1T6, NU214, Q9W204, Q9W254, Q9W2L6, Q9W2V9, Q9W330, Q9W337, Q9W362, Q9W3F3, Q9W3M7, Q9W3V9, Q9W4M7, Q9W4N5, Q9W4X2, Q9W532, Q9W5R5, Q9Y0Z1, Q9Y102, Q9Y1L3, R9TG69, T2FF90, V9GZU6, X2J6N7, X2J6U8, X2J786, X2J847, X2JAC7, X2JAF1, X2JAH8, X2JAZ3, X2JB24, X2JB38, X2JC54, X2JCH4, X2JCM1, X2JCN4, X2JD52, X2JDD7, X2JDJ5, X2JDU0, X2JEB4, X2JEN2, X2JG40, X2JG50, X2JIM3, X2JJG8, X2JL49
Species: Drosophila ananassae, Drosophila melanogaster
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Selvan N, Williamson R, Mariappa D, Campbell DG, Gourlay R, Ferenbach AT, Aristotelous T, Hopkins-Navratilova I, Trost M, van Aalten DMF. A mutant O-GlcNAcase enriches Drosophila developmental regulators. Nature chemical biology 2017 13(8) 28604694
Abstract:
Protein O-GlcNAcylation is a reversible post-translational modification of serines and threonines on nucleocytoplasmic proteins. It is cycled by the enzymes O-GlcNAc transferase (OGT) and O-GlcNAc hydrolase (O-GlcNAcase or OGA). Genetic approaches in model organisms have revealed that protein O-GlcNAcylation is essential for early embryogenesis. The Drosophila melanogaster gene supersex combs (sxc), which encodes OGT, is a polycomb gene, whose null mutants display homeotic transformations and die at the pharate adult stage. However, the identities of the O-GlcNAcylated proteins involved and the underlying mechanisms linking these phenotypes to embryonic development are poorly understood. Identification of O-GlcNAcylated proteins from biological samples is hampered by the low stoichiometry of this modification and by limited enrichment tools. Using a catalytically inactive bacterial O-GlcNAcase mutant as a substrate trap, we have enriched the O-GlcNAc proteome of the developing Drosophila embryo, identifying, among others, known regulators of Hox genes as candidate conveyors of OGT function during embryonic development.
O-GlcNAc proteins:
A0A024QZW7, A0A024R899, A0A0B4KEI6, A0A0B4KFC5, A0A0B4KFJ8, NDG, A6NEM2, A8JV18, B2RAH5, B7Z060, B7Z0D3, D3DUP1, E1JI40, E7EPN9, F8VRH0, M9NE54, ANR17, SC24B, LMNA, ITA2, ATNA, NU214, AGFG1, SC24C, LAMA, UBP2L, CAPR1, NCOA6, SF01, TAB1, ISWI, QSER1, Q59FC9, PRC2B, UBAP2, EGRAP, Q7JQR3, NUP62, NUP50, Q7K3E2, Q7KMM4, Q7KTF8, Q86BH2, Q8INH6, ANKH1, Q8MLT4, PHC3, ATX2, SMAP2, ZFR, ATX2, Q9P0H6, HECD1, SP27A, HCF, Q9V9X0, Q9VAU1, Q9VCD1, Q9VCI4, NUP58, Q9VGL0, Q9VI72, Q9VMM2, Q9VUH6, Q9VW34, Q9VWL7, MINY3, NU153, Q9VXM5, RAE1, Q9W2U7, Q9W3Q5, Q9Y0Z1, S23IP
Species: Drosophila melanogaster, Homo sapiens
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