| 1999 |
ANKFY1 (Ankhzn) protein localizes to endosomal membranes, as determined by immunoelectron microscopy. The protein contains 17 ankyrin repeats hooked to a zinc finger (FYVE) motif, and was confirmed at ~130 kDa by in vitro transcription/translation and antibody detection on SDS-PAGE. |
Immunoelectron microscopy, in vitro transcription/translation, SDS-PAGE/Western blot |
Biochemical and biophysical research communications |
Medium |
10092534
|
| 2000 |
Human ANKFY1 (ANKHZN) protein is present in both membrane and soluble fractions on subcellular fractionation, confirming partial membrane association. The gene maps to chromosome 17p13. |
Subcellular fractionation, Western blot, radiation hybrid panel, FISH |
Gene |
Medium |
10940552
|
| 2003 |
ANKFY1 (Ankhzn) co-localizes with phagocytosed dextran particles and transferrin-labeled endocytotic structures in macrophages, and its immunoreactivity is markedly increased in serum-starved cells, suggesting involvement in endocytosis and autophagy vesicle formation. |
Immunohistochemistry, immunofluorescence co-localization with transferrin and dextran, serum starvation assay |
Kaibogaku zasshi. Journal of anatomy |
Low |
12833855
|
| 2017 |
ANKFY1 acts as a BTB-domain adaptor protein for the CUL3 E3 ubiquitin ligase complex and is required for early endosomal localization of integrin β1 on the cell surface of endothelial cells. CUL3 physically interacts with ANKFY1 and is required for ANKFY1's early endosomal localization. Depletion of either CUL3 or ANKFY1 by siRNA reduces surface integrin β1 levels and impairs angiogenesis. |
siRNA knockdown, co-immunoprecipitation, immunofluorescence/confocal localization, flow cytometry for surface integrin β1, siRNA screen of 175 BTBPs |
Biology open |
High |
29038302
|
| 2018 |
ANKFY1 physically interacts with GAPVD1 (co-immunoprecipitation) and both proteins co-localize in HEK293T cells. Both proteins interact with active RAB5 (GTP-bound form). Patient-derived missense mutations in ANKFY1 alter binding affinity for active RAB5 and reduce ability to rescue knockout-induced podocyte migration defects, implicating ANKFY1 in RAB5-dependent endosomal regulation in podocytes. |
Co-immunoprecipitation, co-localization in HEK293T, siRNA silencing, podocyte migration assay, ectopic expression of patient-derived mutant proteins, Drosophila nephrocyte endocytosis assay |
Journal of the American Society of Nephrology : JASN |
High |
29959197
|
| 2020 |
ANKFY1 knockdown in human retinal microvascular endothelial cells (HRMECs) reduces cell-surface VEGFR2 protein levels (without affecting VEGFR2 mRNA) and attenuates downstream Akt/eNOS signaling, thereby impairing VEGF-dependent and -independent endothelial cell proliferation and migration. |
siRNA knockdown, Western blot for surface VEGFR2 and Akt/eNOS phosphorylation, qRT-PCR, cell proliferation and migration assays |
Biochemical and biophysical research communications |
Medium |
33092793
|
| 2021 |
Ankfy1 knockout mice develop normally but at 9 months show progressive loss of cerebellar Purkinje cells (with other cerebellar cell types largely unaffected) and defective motor function, establishing a cell-type-specific role for ANKFY1 in Purkinje cell maintenance in vivo. |
Whole-body knockout mouse model, histology, immunofluorescence, motor behavior tests |
Frontiers in cellular neuroscience |
Medium |
33796010
|
| 2024 |
ANKFY1 is an endosome-localized protein that binds PI3P through its FYVE domain, directly interacts with ATG2A, and promotes ATG2A-mediated lipid transfer from PI3P-containing liposomes. A pool of ANKFY1 co-localizes with ATG2A between endosomes and phagophores. Depletion of ANKFY1 impairs autophagosome growth and reduces autophagy flux, phenocopying ATG2A/B depletion. Depletion of UVRAG, ANKFY1, or ATG2A/B reduces PI3P on phagophores, placing ANKFY1 in the UVRAG–PI3P–ATG2A lipid transfer pathway from endosomes to phagophores. |
Co-immunoprecipitation/pulldown, in vitro lipid transfer assay with purified recombinant proteins and PI3P-containing liposomes, siRNA/shRNA depletion, autophagy flux assays, confocal co-localization, genetic epistasis (UVRAG/ANKFY1/ATG2A/B depletion) |
Cell discovery |
High |
38622126
|
| 2016 |
Ankfy1 is dispensable for neural stem/precursor cell development in mice with a mixed genetic background, but Ankfy1 knockout is lethal by embryonic day 11.5 in a pure C57BL/6 inbred background, indicating an essential but genetically background-dependent role in early embryonic development. |
Knockout mouse generation, immunofluorescence, in situ hybridization, genotyping by PCR |
Neural regeneration research |
Medium |
28123425
|
| 2024 |
Compound heterozygous ANKFY1 variants (p.Ser918Ter and a splice-site deletion) found in a patient with proteinuria and movement disorder lead to reduced ANKFY1 protein expression in vitro, supporting a loss-of-function mechanism for bi-allelic ANKFY1 variants in a neuro-renal syndrome. |
Whole-exome sequencing, in vitro functional study (Western blot for protein expression of patient variants) |
Clinical kidney journal |
Low |
38915441
|
| 2026 |
ANKFY1 interacts with PDCoV nonstructural protein nsp8, recruits the E3 ubiquitin ligase Cullin3 to catalyze K63/K33-linked ubiquitination of nsp8 at lysine 58, and the ubiquitinated nsp8 is subsequently recognized by selective autophagy receptor p62 and delivered to autolysosomes for degradation, thereby suppressing viral replication. |
Co-immunoprecipitation, gain/loss-of-function (overexpression and siRNA depletion), ubiquitination assay with site-directed mutagenesis (K58 site), p62 interaction assay, viral replication and RNA synthesis quantification |
Veterinary microbiology |
Medium |
41763084
|
| 2023 |
miR-760 directly targets the ANKFY1 3'UTR (validated by luciferase reporter assay) and suppresses ANKFY1 protein expression. ANKFY1 participates in APS-mediated promotion of osteogenic differentiation and proliferation of human bone marrow mesenchymal stem cells. |
Luciferase reporter assay, Western blot, qRT-PCR, miR-760 overexpression/knockdown, osteogenic differentiation assays |
Bone & joint research |
Low |
37532241
|