Annotated protein:Liprin-alpha-3 (Protein tyrosine phosphatase receptor type f polypeptide-interacting protein alpha-3) (PTPRF-interacting protein alpha-3). Gene symbol: PPFIA3. Taxonomy: Mus musculus (Mouse). Uniprot ID: P60469
antibody wiki:
SynGO gene info:SynGO data @ PPFIA3
Ontology domain:Cellular Component
SynGO term:presynaptic active zone (GO:0048786)
Synapse type(s):hippocampus, glutamatergic
Annotated paper:Wong MY, et al. "Liprin-alpha3 controls vesicle docking and exocytosis at the active zone of hippocampal synapses" Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):2234-2239 PMID:29439199
Figure(s):Fig. 1, Fig. S1
Annotation description:Cultured hippocampal neurons stained with antibodies against the synaptic vesicle markers - vGlut or synapsin, the active zone protein -Bassoon, and liprin alpha 3 were imaged using STED microscopy and pairwise stainings of synapses in side view were analyzed for all known AZ proteins and liprin alpha 3. To assess the localization of each protein within and outside the active zone, two masks were generated (Fig. 1A): one that outlined the nerve terminal (vGlut1 or Synapsin; Fig. 1A, blue), and one that defined the active zone (BassoonN; Fig. 1A, green). The area associated with the active zone for each protein was quantified (Fig. 1A, yellow) and compared with the area of the same protein outside the active zone (Fig. 1A, red), normalized to the sum of the area of active zone and nonactive zone components (Fig. 1E). This provides a measure of signal distribution within a nerve terminal without accounting for the signal intensity within the area. For Bassoon, RIM1, ELKS2, RIM-BP2, and Munc13-1, the majority of the area stained for was at the active zone (73-90%; Fig. 1E). For Liprin-α3, ~50% of the positively stained area was outside the active zone but the peak intensity was within the active zone.

20/01/2021 Dnyanada: Information on antibody specificity:
Figure 2C: Western blot demonstrates the absence of anti-Liprin-α3 signal in lysates derived from Liprin-α3-KO mice. However, anti-Liprin-α3 cross reacts with Liprinα1-4.
Evidence tracking, Biological System:Intact tissue
Cultured neurons
Evidence tracking, Protein Targeting:Antibody (detection)
Evidence tracking, Experiment Assay:Super resolution
Confocal
Annotator(s):Arthur de Jong (ORCID:0000-0002-7620-2704)
Pascal Kaeser (ORCID:0000-0002-1558-1958)
Lab:Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA
Additional literature:Thus study supports synaptic localization of liprin alpha 3 using confocal microscopy.

Figure 6: Immunohistochemical costaining of Liprin-a3 and the presynaptic marker synapsin on sagittal adult mouse cerebellar sections demonstrates co-labeling.

Figure 11: Confocal microscopy images of immunocytochemistry of primary mouse cortical neurons DIV14-21. Liprin-alpha 3 shows strong co-localization with the presynaptic protein Bassoon.

Liprin alpha 3 antibodies were raised in rabbits against the peptide: RGRPPSSYSRSLPGSC, affinity purified, and used at 1:20 (ICC) and 1:50 (IHC). @ PMID:21618221

Thus study supports synaptic localization of liprin alpha 3 using confocal microscopy.

Figure 6: Immunohistochemical costaining of Liprin-a3 and the presynaptic marker synaptophysin in the in mossy fiber endings in the stratum lucidum of the hippocampus of adult mouse brain sections demonstrates co-labeling.

Figure 9: Confocal immunofluorescence imaging of Liprin-a3 and the presynaptic marker bassoon at synapses in hippocampal neuronal cultures demonstrates co-labeling.

Rabbit polyclonal anti-liprin-a antibodies were raised
against GST-tag fusion proteins containing sequences that showed the lowest amount of homology with other lipton alpha family members (187-286 of liprin-a3 (NM_003660.2)).
Rabbit anti-liprin-a3 was used at-1:100 for ICC, 1:2000 for IHC, and 1:500 for IF. @ PMID:21618222
SynGO annotation ID:4171
Dataset release (version):20231201
View annotation as GO-CAM model:Gene Ontology