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ProteomicsEngineering Af1521 macro domain increases ADP-ribose binding affinity and leads to the identification of more ADP-ribosylated proteins
Protein ADP-ribosylation is a covalent, reversible post-translational modification that has important functions in several cellular mechanisms. The identification of modified proteins in cells has proven challenging and, due to the low abundance of protein ADP-ribosylation, has mainly been possible ...
ORGANISM(S): Homo sapiens (Human) 
2020-10-20 | PXD016686 | Pride
Ap-ms Af1521 macro domain Adp-ribosylation Lc-ms/ms 
ProteomicsAffinity purification of ADP-ribosylated proteins from A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1 (2.5 ug Af1521 Macro, technical replicate 1)
ADP-ribosylated proteins were enriched using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) from transgenic A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1. Enriched proteins were analyzed by LC-MS/MS.
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2023-09-28 | PXD045544 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation 
ProteomicsAffinity purification of ADP-ribosylated proteins from A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1 (5 ug Af1521 Macro, technical replicate 2)
ADP-ribosylated proteins were enriched using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) from transgenic A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1. Enriched proteins were analyzed by LC-MS/MS.
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2023-09-28 | PXD045545 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation 
ProteomicsAffinity purification of ADP-ribosylated proteins from A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1, DDA method (2.5 ug Af1521 Macro, technical replicate 1)
ADP-ribosylated proteins were enriched using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) from transgenic A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1. Enriched proteins were analyzed by LC-MS/MS.
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2023-09-28 | PXD045558 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation 
ProteomicsAffinity purification of ADP-ribosylated proteins from A. thaliana plants expressing the Pseudomonas syringae type III effector AvrRpm1, DDA method (5 ug Af1521 Macro, technical replicate 2)
ADP-ribosylated proteins were enriched using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) from transgenic A. thaliana lines expressing the Pseudomonas syringae type III effector AvrRpm1. Enriched proteins were analyzed by LC-MS/MS.
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2023-09-28 | PXD045548 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation 
ProteomicsSupplementary Dataset S7. ADP-ribosylated peptides identified in two Af1521 Macro pulldown experiments from HopU1-Flag vs. Col-0 samples by the ‘triggering’ method.
During infection, the plant pathogenic bacterium Pseudomonas syringae translocates type 3 effector proteins into the host cell. The effector protein HopU1 is an ADP-ribosyltransferases that ADP-ribosylates host proteins to subvert plant innate immunity. The aim of this experiment was to compare affi...
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2025-11-17 | PXD068739 | Pride
Hopu1 Lc-msms Arabidopsis thaliana Affinity purification Plant Pseudomonas syringae Adp-ribosylation Plant pathogen Type 3 effector 
ProteomicsComparison of proteins identified using the Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) versus the non-functional (G42E) Macro domain for enrichment of ADP-ribosylated proteins from A. thaliana plants expressing the Pseudomonas syringae type III effectors AvrRpm1 or HopF2 (DDA method)
ADP-ribosylated proteins were enriched using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) from transgenic A. thaliana plants expressing the Pseudomonas syringae type III effectors AvrRpm1 or HopF2. A non-functional Af1521G42E mutant of the engineered ...
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2023-09-28 | PXD045726 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation 
ProteomicsSupplementary Dataset S4. Comparison of proteins identified in three experiments using the functional (Af1521) versus non-functional (G42E) Macro domains following ADPr-enrichment of proteins from Dex-treated AvrRpm1-HA plants.
During infection, the plant pathogenic bacterium Pseudomonas syringae translocates type 3 effector proteins into the host cell. The effector protein AvrRpm1 is an ADP-ribosyltransferases that ADP-ribosylates host proteins to subvert plant innate immunity. The aim of this experiment was to compare af...
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2025-11-17 | PXD068551 | Pride
Lc-msms Arabidopsis thaliana Affinity purification Plant Pseudomonas syringae Adp-ribosylation Avrrpm1 Plant pathogen Type 3 effector 
ProteomicsAffinity purification of ADP-ribosylated proteins from A. thaliana plants expressing the Pseudomonas syringae type III effector HopU1
ADP-ribosylated proteins were enriched using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w) from transgenic A. thaliana lines expressing the Pseudomonas syringae type III effector HopU1. A non-functional Af1521 G42E mutant of the engineered Macro domain...
ORGANISM(S): Arabidopsis thaliana (Mouse-ear cress) 
2024-10-22 | PXD056608 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation 
ProteomicsPseudomonas type III effector AvrRpm1 ADP-ribosylates Arabidopsis thaliana MEMBRANE-ASSOCIATED MANNITOL-INDUCED (MAMI)
Nicotiana benthamiana tissue expressing either AvrRpm1-3xFlag, StrepII-3xHA-MAMI, or co-expressing AvrRpm1-3xFlag and StrepII-3xHA-MAMI were subjected to enrichment of ADP-ribosylated proteins using the engineered Af1521 Macro domain (Nowak et al., https://doi.org/10.1038/s41467-020-18981-w). A non-...
ORGANISM(S): Nicotiana benthamiana 
2023-09-28 | PXD045549 | Pride
Arabidopsis thaliana Type iii effector Pseudomonas syringae Plant pathogen Adp-ribosylation Nicotiana bethamiana 
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