GAD67 is a PLP-dependent glutamate decarboxylase that supplies GABA. The active full-length enzyme, its inactive GAD25 splice product, engineered structures and interaction-screen clones must remain distinct. Current judgments and manual reference assessments are in the review; literal annotation contexts and historical locators are in the source archive.
The original GAD67/GAD25 comparison measures radiolabeled glutamate-to-CO2 activity using equimolar translation, PLP and controls. Full-length GAD67 is active and GAD25 inactive; the measured reaction directly supports both L-glutamate breakdown and GABA biosynthesis within one catalytic core; the variant-focused title does not negate canonical activity (PMID:10671565(https://pubmed.ncbi.nlm.nih.gov/10671565/)). Cloned human GAD67 is independently enzymatically active in bacterial expression (PMID:1549570(https://pubmed.ncbi.nlm.nih.gov/1549570/)). The GAD1 structure and catalytic analysis distinguish sustained GAD67 activity from GAD65 regulation (PMID:17384644(https://pubmed.ncbi.nlm.nih.gov/17384644/)).
Official PDB 2OKJ is human GAD1, with a homodimeric biological assembly. Its engineered entity is canonical residues 93–594 plus HH, 504 residues in total. It is not a natural splice product; 2OKK is the separate GAD2 structure. Complete subscription Methods were not independently obtained.
| Current product | Sequence scope | Functional interpretation |
|---|---|---|
| Q99259-1 / GAD67 | Displayed, 594 residues; direct FASTA exact | Characterized active full-length enzyme |
| Q99259-2 | UniProt says not described; no sequence recovered | No invented FASTA or activity assignment |
| Q99259-3 / GAD25 | 224 residues; direct FASTA exact | Original GAD25 evidence establishes inactivity |
| Q99259-4 | 425 residues; direct FASTA exact | Sequence verified; equivalent catalysis/targeting not assumed |
The isoform-interaction study’s GAD1_1 clone translates exactly to the current 594-residue reference. Study GAD1_2 and GAD1_3 are experimental clone names, not automatically current UniProt suffixes (PMID:26871637(https://pubmed.ncbi.nlm.nih.gov/26871637/)).
The SH3 screen places the GAD1 peptide at residues 520–532 opposite NCK1 SH3 domains; GAD1 is the domain-binding partner, not the SH3-domain provider (PMID:17474147(https://pubmed.ncbi.nlm.nih.gov/17474147/), original Table S2). The other positive binary screens remain documented with exact partners, suffixes and construct limits. Their generic protein-binding annotations are removed as functionally uninformative; removal does not deny the reported interactions, and no unsupported specific molecular function is substituted. HuRI explicitly tests Q99259-1; its array, pooling and validated records are related assay forms, not three independent physiological replications (PMID:32296183(https://pubmed.ncbi.nlm.nih.gov/32296183/)). HAAO T42S/I37V belong to the partner in the variant screen, not to GAD1 (PMID:31515488(https://pubmed.ncbi.nlm.nih.gov/31515488/), Supplementary Data 2 rows 476–477).
One source join remains unresolved: the complete 1,166-pair Table 2 of PMID:21516116(https://pubmed.ncbi.nlm.nih.gov/21516116/) pairs GAD1/2571 with CMTM5/116173, while the incoming annotation names AGTRAP/57085. Other papers independently support GAD1–AGTRAP, so failure to recover this source-specific pair is not evidence that binding is false. Likewise, the selected GAD25-paper Discussion does not independently resolve its GAD1–GAD2 IPI assay.
Actual GAD67 antibody evidence supports cytoplasm and inhibitory presynaptic terminals (PMID:15347806(https://pubmed.ncbi.nlm.nih.gov/15347806/), selected Results/Figure 2; PMID:23258346(https://pubmed.ncbi.nlm.nih.gov/23258346/), antibody Methods/Figure 3; PMID:24613359(https://pubmed.ncbi.nlm.nih.gov/24613359/), selected Results/Figures 1 and 4). This is distinguished from GFP-at-Gad1-locus cell identification in the accessible PMID:21584649(https://pubmed.ncbi.nlm.nih.gov/21584649/) abstract. Its exact source assay remains uninspected. The source paper has no verified PMC identifier; an earlier citation-list extraction was corrected rather than treated as a retraction or remapping.
Actual PAINT PTHR45677 assertions and the exact 594-residue target alignment support all four incoming ancestral placements. The catalytic/GABA-synthesis node is PTN002903772, cytoplasm PTN000242648 and active zone PTN002903810. Current versus literal/main donor differences remain separately dated. The active-zone original panels in PMID:12074840 remain inaccessible; the annotation retains the actual curator/PAINT placement without invented antibody or ultrastructure details. Selected PMID:18165320 electron microscopy labels SLC4A10, not GAD67. Precise perinuclear and cell-cortex claims remain unresolved.
The official GAD1 component R-HSA-888580 is human Q99259, chain 1–594, at the GABA transport-vesicle membrane. The GAD1 synthesis reaction includes its homodimer; the GAD2-labelled reaction includes a GAD1:GAD2 assembly. Release and docking contain the same target through vesicle assemblies. Thus pathway labels do not establish target absence. Conversely, the release summary locates SNAP25 and syntaxin at the plasma membrane, while its GAD1 component remains on the transport vesicle. The GAD1 plasma-membrane TAS is therefore marked over-annotated for this specific event-to-participant inference, without a universal claim of cell-surface exclusion. Both the broad vesicle-membrane row and the four specialized transport-vesicle rows are kept non-core for this same pool. The homodimer chain means recruitment is not asserted to occur exclusively through GAD1:GAD2 heterodimers. The model is not a new demonstration of GAD1 fusion/docking activity or equal targeting of every product.
The GABA shunt explicitly starts with glutamate decarboxylation, so GAD1 does perform its entry step; downstream reactions and complete physiological flux are not thereby established. Chemical synaptic transmission specifically concerns release and postsynaptic activation. Supplying GABA alone does not resolve the separate source-specific transmission claim.
The inherited review contains 31 source rows and a separate cytosol NEW. Its complete immutable Git locator, all 37 quotation-association locators and the original journal prefix are retained in the archive. Seven short excerpts now anchor the current review: the inherited basal-GABA statement, the completed bacterial-expression sentence and five additional exact cached reaction, cofactor, compartment, PDB and SUBUNIT records. The original 37 associations remain accessible in the linked historical review, including their literal spellings. The old cytosol NEW is not reasserted: the actual IBD is cytoplasm, and its family-membership quote does not establish the narrower compartment. No absence of biological cytosol localization is claimed.
Normal cached text and separately accessed originals are described distinctly in each reference assessment. Partial Introduction/Discussion caches, authentic selected indexed originals and inaccessible source-specific panels are not called complete Methods reads. Genuine native acquisition reconciled all 50 original contexts with the actual 16-column export and private native seed. All 46 source files and the two initial inherited quotations were checked against their actual bytes; 21 main sources were reused unchanged and 25 missing sources were acquired. Cache full-text availability is recorded separately from the exact sections assessed. The inherited review and journal remained untouched throughout acquisition. No current decision is duplicated in the compact evidence archive.
The current excerpts are in the review rather than duplicated here. They can be checked directly against the repository's human-expression abstract, GAD65/GAD67 catalytic abstract, GAD1 reaction, GAD1 vesicle component and UniProt PDB and SUBUNIT statements. The PDB line identifies the crystallized chain range; the SUBUNIT statement directly names the homodimer with experimental evidence from PMID:17384644, corroborated by the separate 2OKJ assembly record. A reaction title or compartment label is model evidence, not an independently performed localization assay.
For externally accessed supplements, the source locators remain claim-specific: PMID:17474147(https://pubmed.ncbi.nlm.nih.gov/17474147/) Table S2 (NCK1 domains and GAD1 peptide); PMID:31515488(https://pubmed.ncbi.nlm.nih.gov/31515488/) Supplementary Data 2 rows 476–477 (HAAO variants); and PMID:25416956(https://pubmed.ncbi.nlm.nih.gov/25416956/) Supplementary Table 2G rows 1854–1857 (binary pairs). These selected original records are not represented as text inside the abstract-only normal caches. The complete inherited review at its immutable Git revision exposes all historical wording without requiring access to private files.
The five remaining unresolved judgments are unchanged. For PMID:21516116, 8954991 and 29960053, the identifier is established but the exact annotation-to-experiment support has not been verified. The available excerpts or missing pair join do not demonstrate a wrong PMID or biological contradiction.