| 2002 |
X-ray crystal structure of the GGA1 VHS domain alone and in complex with the cation-independent mannose 6-phosphate receptor (CI-MPR) C-terminal peptide containing an acidic-cluster dileucine (ACLL) sequence revealed that the VHS domain forms a super-helix of eight alpha-helices; recognition of ACLL motifs involves unidirectional movements of helices α6 and α8 creating electrostatic and hydrophobic interactions. |
X-ray crystallography (crystal structures of apo and peptide-bound VHS domain) |
Nature |
High |
11859376
|
| 2002 |
Full-length cytoplasmic GGA1 (and GGA3 but not GGA2) is autoinhibited in its ability to bind the CI-MPR because an acidic-cluster dileucine (AC-LL) sequence in the hinge segment binds to the VHS domain ligand-binding site; this autoinhibition depends on phosphorylation of a serine three residues upstream of the AC-LL motif by casein kinase 2. |
In vitro binding assays with full-length and truncated GGA constructs, site-directed mutagenesis, in vitro casein kinase 2 phosphorylation assay, substitution of GGA1 inhibitory sequence into GGA2 |
Proceedings of the National Academy of Sciences of the United States of America |
High |
12060753
|
| 2002 |
The VHS domains of GGA1 and GGA2 bind the cytosolic domain of memapsin 2 (beta-secretase/BACE); Asp496, Leu499, and Leu500 in the memapsin 2 cytosolic tail are essential for this interaction, mirroring the spacing found in mannose-6-phosphate receptor cytosolic domains. |
Gel-immobilized VHS domain pulldown from cell lysates, site-directed mutagenesis of memapsin 2 cytosolic tail |
FEBS letters |
Medium |
12135764
|
| 2002 |
The cytoplasmic domain of the Vps10p-domain receptor sorLA binds GGA1 and GGA2 via three critical C-terminal residues that conform to a minimal GGA-binding motif (ψ-ψ-X-X-φ) that lacks a classical acidic cluster or dileucine. |
In vitro binding assays, mutagenesis of sorLA cytoplasmic tail |
FEBS letters |
Medium |
11821067
|
| 2003 |
Live-cell fluorescence imaging showed that GGA1, clathrin, and AP-1 colocalize at the TGN and on pleomorphic vesicular-tubular carriers (up to 10 µm displacement at ~1 µm/s) that bud from the TGN; GGA1 and clathrin cycle on and off membranes with half-times of 10–20 s independently of vesicle budding. |
Live fluorescence imaging with GFP-tagged GGA1, clathrin, and AP-1; FRAP |
Molecular biology of the cell |
High |
12686608
|
| 2003 |
Crystal structure of the GGA1 GAT domain at 2.4 Å revealed a three-helix bundle with a long N-terminal helical extension; the ARF-binding site resides in the N-terminal extension, and the core three-helix bundle shares structural homology with the N-terminal domain of syntaxin 1a. |
X-ray crystallography at 2.4 Å resolution |
Proceedings of the National Academy of Sciences of the United States of America |
High |
12668765
|
| 2003 |
The 2.8 Å crystal structure of the GGA1 GAT domain revealed four helices; the conserved N-terminal helix-loop-helix motif harbors a hydrophobic ARF-binding patch, and the C-terminal three-helix bundle is responsible for rabaptin-5 binding, as confirmed by structure-based mutagenesis. |
X-ray crystallography at 2.8 Å, structure-based mutagenesis and biochemical binding assays |
Biochemistry |
High |
12767220
|
| 2003 |
The C-terminal three-helix bundle of the GGA1 GAT domain mediates binding to the coiled-coil region of Rabaptin-5 through a hydrophobic surface patch; the GGA1-specific residue N284 (vs. S293 in GGA3) accounts for the differential Rabaptin-5 binding among GGA family members, and GAT–Rabaptin-5 binding is independent of ARF binding. |
Site-directed mutagenesis, in vitro pulldown/binding assays, crystal-structure-guided interpretation |
Biochemistry |
High |
14636058
|
| 2004 |
The BACE cytosolic DISLL motif is recognized by the GGA1 VHS domain; phosphorylation of the serine in this motif enhances binding affinity ~3-fold. The crystal structure of the GGA1 VHS domain bound to the phosphorylated BACE peptide showed that phospho-serine alters the lysine side chain and backbone, creating an additional hydrogen bond and stronger electrostatic interaction. |
X-ray crystallography of VHS–BACE peptide complex, quantitative binding assays with phosphorylated and unphosphorylated peptides |
Traffic (Copenhagen, Denmark) |
High |
15117318
|
| 2004 |
Fluorescence lifetime imaging microscopy (FLIM/FRET) in intact cells demonstrated that GGA1 interacts with phosphorylated BACE in juxtanuclear compartments; serine phosphorylation of BACE regulates its interaction with GGA1 in cells, and non-phosphorylatable or pseudo-phosphorylated BACE mutants remain colocalized with GGA1 at the Golgi. |
FLIM-FRET in live/fixed cells, phosphomimetic and non-phosphorylatable BACE mutants |
Journal of cell science |
High |
15466887
|
| 2004 |
GGA1 interacts with the AP-1 gamma-ear through a WNSF sequence (W382-N383-S384-F385) in its hinge region; Trp and Phe are critical residues; this WXXF-type motif competes with Rabaptin-5 FXXPhi-type binding for the same or overlapping site on the AP-1 gamma-ear. |
In vitro binding assays, competitive inhibition with peptides, site-directed mutagenesis of GGA1 hinge and AP-1 gamma-ear |
The Journal of biological chemistry |
High |
14973137
|
| 2004 |
Glu58 and Glu59 of the CD-MPR CK2 acidic cluster site are essential for high-affinity GGA1 binding in vitro; phosphorylation of Ser57 does not influence GGA1 binding to CD-MPR. The binding affinity of GGA1 to CD-MPR is 2.4-fold higher than that of AP-1. |
In vitro binding assays with phosphopeptides and mutant CD-MPR cytoplasmic tail peptides, quantitative affinity measurements |
The Journal of biological chemistry |
Medium |
15044437
|
| 2004 |
GGA1 phosphorylation at S268 and T270 in the GAT domain (identified by tandem mass spectrometry) causes redistribution from Golgi/TGN to cytoplasmic puncta when phosphomimetic mutations are introduced; this phosphorylation regulates the rate of GGA1 coat dissociation from vesicles. |
Tandem mass spectrometry to identify phosphorylation sites, expression of phosphomimetic HA-GGA1 mutants in mammalian cells, quantitative colocalization with Golgi/TGN markers |
Traffic (Copenhagen, Denmark) |
Medium |
14690499
|
| 2006 |
GGA1 overexpression in cultured cells increased APP C-terminal fragment from beta-cleavage but reduced Abeta production; FRET analysis showed GGA1 confines APP to the Golgi where the two proteins come into close proximity; the GAT domain integrity is required for these effects, and direct GGA1–BACE binding is not required. |
Overexpression and dominant-negative constructs, FRET, subcellular fractionation, domain deletion/mutation analysis |
The Journal of neuroscience |
Medium |
17005855
|
| 2006 |
GGA1 overexpression reduces Abeta secretion while RNAi-mediated knockdown increases Abeta secretion; GGA1 modulates APP processing by affecting subcellular trafficking of BACE1, independent of direct GGA1–APP interaction, and without altering total cellular BACE1 activity. |
GGA1 overexpression and siRNA knockdown in cultured cells, Abeta ELISA, APP processing assays |
The Journal of neuroscience |
Medium |
17151287
|
| 2007 |
GGA1 promotes assembly of clathrin in vitro; full-length GGA1 polymerizes clathrin into both baskets and tubules (~180 nm long, ~50 nm wide); the hinge+GAE fragment assembles clathrin only into baskets; maximum clathrin assembly occurs at one GGA1 per heavy chain. |
In vitro clathrin assembly assay with purified components, electron microscopy to determine structure of assembled complexes |
The Journal of biological chemistry |
High |
17344219
|
| 2007 |
The GGA1 GAE domain crystal structure in complex with its own hinge WNSF peptide revealed that the two aromatic residues fit into a hydrophobic groove of the GAE domain; the hinge region competes with accessory proteins and AP-1 for GAE binding, establishing an autoregulatory mechanism for GGA1 in clathrin-mediated trafficking. |
X-ray crystallography of GAE–hinge peptide complex, fluorescence quenching competition assays |
Traffic (Copenhagen, Denmark) |
High |
17506864
|
| 2012 |
GGA1 silencing potentiates BACE1 elevation induced by GGA3 deletion in neurons in vitro, indicating that GGA1 and GGA3 synergistically regulate BACE1 levels; GGA1 is a caspase-3 substrate that is depleted after traumatic brain injury. |
siRNA knockdown in primary neurons, caspase-3 cleavage assays, mouse TBI model, immunoblotting |
The Journal of neuroscience |
Medium |
22836275
|
| 2012 |
siRNA knockdown of GGA1 (but not GGA2 or GGA3) disrupts LR11/SorLA endosomal trafficking and prevents LR11-mediated and BACE1-mediated modulation of APP processing to Abeta; GGA1 is specifically required for LR11 endocytic traffic. |
siRNA knockdown of individual GGA family members, APP processing assays, BACE1 mutagenesis (S498A), subcellular localization by immunofluorescence |
Molecular biology of the cell |
Medium |
22621900
|
| 2014 |
GGA1 functions in a Rabep1/GGA1/Arl3-dependent ciliary targeting mechanism at the TGN; GGA1 couples the polycystin-1/polycystin-2 complex (identified by yeast two-hybrid) to an Arl3-based ciliary trafficking module, enabling trafficking of these large membrane proteins to cilia. |
Yeast two-hybrid screening, candidate approach, co-immunoprecipitation, siRNA knockdown with ciliary localization readout |
Nature communications |
Medium |
25405894
|
| 2017 |
GGA1 mediates rapid trafficking of phosphorylated BACE1 (phospho-S498, DISLL motif) from early endosomes to recycling endosomes; the phosphomimetic S498D mutant exits early endosomes faster and shows reduced APP processing and Abeta production; retromer cooperates with GGA1 in this pathway. |
Phosphomimetic/non-phosphorylatable BACE1 mutants, siRNA knockdown of GGA1 and retromer, quantitative endosomal trafficking assays, Abeta ELISA, primary neuron experiments |
Molecular biology of the cell |
High |
29142073
|
| 2016 |
GGA1 and GGA2 are required for cell surface transport of α2B-adrenergic receptor (α2B-AR); knockdown of GGA1 arrests α2B-AR in the perinuclear region and attenuates ERK1/2 activation and cAMP inhibition; the GGA1 hinge region directly interacts with the third intracellular loop of α2B-AR. |
shRNA/siRNA knockdown, cell surface ELISA, receptor-mediated signaling assays, co-immunoprecipitation, domain mapping |
Scientific reports |
Medium |
27901063
|
| 2019 |
A naturally occurring truncated splice variant of GGA1 (GGA1t), lacking the N-terminal hinge portion, acts as a dominant-negative inhibitor of α2B-AR cell surface export; GGA1t forms homodimers and heterodimers with full-length GGA1, is unable to bind cargo α2B-AR, and cannot recruit clathrin to the TGN. |
Overexpression of GGA1t, cell surface receptor assays, Co-IP for dimerization, clathrin recruitment assay at TGN |
Scientific reports |
Medium |
31316103
|
| 2018 |
GGA1 is required for myotube formation in C2C12 myoblasts; Gga1 depletion by RNAi prevents formation of large multi-nucleated myotubes; GGA1 is involved in cell surface expression and sorting of the insulin receptor, as inhibition of lysosomal proteases in GGA1-knockdown cells increased insulin receptor levels. |
siRNA knockdown in C2C12 cells, morphological analysis of myotube formation, lysosomal protease inhibition, insulin receptor immunoblotting |
PloS one |
Medium |
30440034
|
| 2024 |
GGA1 interacts with the endosomal Na+/H+ exchanger NHE6 (and organellar NHEs 6, 7, 9 but not surface NHEs 1 and 5) via the NHE6 cytoplasmic tail; GGA1 knockout causes NHE6 mislocalization — less NHE6 in endosomes, more in lysosomes and Golgi, with increased surface exocytosis — and alkalinization of Golgi luminal pH. |
Yeast two-hybrid screening, reciprocal co-immunoprecipitation (overexpressed and endogenous), hybrid NHE1/NHE6 domain-swap constructs, subcellular fractionation in GGA1 KO cells, super-resolution microscopy co-localization, luminal pH measurement |
The Journal of biological chemistry |
High |
39002678
|
| 2025 |
In GIST cells, a constitutively active KIT mutant activates the PLCγ2–PKD2–PLD2 cascade, which promotes association of γ-adaptin with GGA1 at the Golgi/TGN; PLD activity is required for GGA1-dependent Golgi/TGN retention of KIT mutant, and PLD inhibition releases KIT mutant from the Golgi for lysosomal degradation. |
PLD inhibitor treatment, siRNA knockdown of PLD1/PLD2, co-immunoprecipitation of γ-adaptin with GGA1, subcellular localization assays |
bioRxivpreprint |
Low |
bio_10.1101_2025.03.02.640696
|
| 2005 |
In vitro biochemical characterization combined with the GAT–Rabaptin-5 complex crystal structure established that the binding mode between the GGA1 GAT domain and Rabaptin-5 is helix-bundle-to-helix-bundle in nature. |
In vitro binding assays, crystal structure of GGA1 GAT–Rabaptin-5 complex |
Methods in enzymology |
Medium |
16473621
|