The sections below identify the original experiment, its public access route and its limitations. Original machine records and publication caches are unchanged. No new publication quotations are supplied. The primary-source assessment below is complemented by the database and donor assessment later in this guide.
Original record; inspected access route.
Actual author-upload original: Results on enzyme activity and Figures 2B,3–8; Methods cloning, bacterial GST preparation,32P-myelin-basic-protein assay and cell fractionation were read. Human fetal-brain 331-aa LDH1, untagged/Myc-His/EGFP HeLa expression and separate bacterial GST enzyme are distinguished. PKA/c-Abl labeling supports Ser/Thr and Tyr substrate capacity after 8 h incubation; this is not endogenous substrate identification. Polysome-associated immunogold/fractionation does not establish a direct ER-membrane interaction. Figure captions were inspected, not image pixels.
Original record; inspected access route.
PubMed/normal-cache abstract read; it explicitly reports plasma-membrane/ER location and tyrosine phosphatase activity. The indexed original PDF mirror exposed title/opening text but timed out on complete retrieval; no operative original or image was inspected. Independent PMID:11001928 resolves broad protein phosphatase and cytoplasmic claims; membrane attachment remains unresolved. No title-based rejection.
Original record; inspected access route.
Actual author-upload Experimental Procedures and Results/Figures 1–4 were read, including protein-free glycogen binding, no-glycogen aggregation control, CBD mutants and amylase release. Human-muscle 331-aa pET21a-His6/FLAG/EGFP preparations are explicit; COS1/HEK293 are hosts. The phosphatase assay is pNPP and does not itself establish protein-tyrosine specificity. Main captions were read, no pixels or separate supplement.
Original record; inspected access route.
Normal/PubMed abstract identifies C-terISO as nuclear plus cytoplasmic and describes unchanged polysome affinity. Complete Methods/Results were not recovered after bounded owner and ROOT publisher, exact-title/PDF and author-page searches. The source is retained; individual splice-product localization is not reconstructed from the abstract. Independent PMIDs:17908927/22961547 supports a context-dependent nuclear pool, not identical localization of all products.
Original record; inspected access route.
The normal/PubMed abstract reports HIRIP5 association and in-vitro dephosphorylation. Publisher and author routes did not provide full text; the indexed CiteSeer PDF failed. No operative HIRIP5 experiment is claimed as inspected. General protein-phosphatase capacity is independently supported by PMID:11001928, while the source-specific interaction remains UNDECIDED.
Original record; inspected access route.
Normal/PubMed abstract and official institutional article metadata were read. PublisherPDF returned403; the UCM repository provides a file record explicitly marked restricted, and attempted download timed out. No original Methods/Results are claimed. The later actual PMID:18070875 endogenous human association and reconstitution resolve PTG adaptor function independently.
Original record; inspected access route.
Normal/PubMed abstract read. It reports a mutation limited to the cytoplasmic isoform, reduced phosphatase activity and retained ER targeting. Complete operative original remains unavailable. No mutation/product assay details beyond the abstract are invented. Independent human experiments resolve broad cytoplasmic/nuclear location; the membrane-specific source context remains unresolved.
Original record; inspected access route.
Actual author-upload Results Generation, brain/liver localization, native human biopsy and glucan-binding sections; main captionsFigures 1–7 and Methods transgene generation were read. The transgene is explicitly human C266S myc-laforin in mice with approximately 100-fold expression; endogenous human EPM2B-mutant biopsy and recombinant human GST-laforin binding are separate. Soma/dendrite/near ER signal is not an endogenous wild-type human localization assay; near ER signal may reside on polyglucosan. Purified protein binds starch more strongly than glycogen. No image pixels inspected.
Original record; inspected access route.
Actual PMC main Methods 11–18, Results 19–40 and Discussion 41–49, all main captions, read. Human malin plus laforin constructs tracing to11739371; human-brain-library two-hybrid, recombinant laforin/in-vitro-translated malin binding and HEK293T co-IP distinguish association from subsequent malin-catalyzed ubiquitination. Laforin C266S ubiquitination does not require its catalysis. Detailed supplementary protocols/pixels were not read.
Original record; inspected access route.
Owner and independentROOT read actual PMC Methods/Results and Figures1–5 captions. WT versus C266S releasesPi from potato amylopectin; glycogen release was not detected in this assay. Negative candidate-protein experiments do not exclude every protein substrate. Construct lineage refers to11739371 human 331-aa. No supplementary or pixel inspection.
Original record; inspected access route.
Actual PMC Results 22–26/Figure 3C and Methods 55–67 read. FLAG-laforin in human HepG2 cells is cytoplasmic at baseline and partly nuclear under glucose deprivation/FSK/IBMX; mutantC266S result is reported in text only. Construct references trace to15930137, while mouse-liverAGL is separate. Only the pertinent localization scope was read; no claim of complete article/supplement/pixel review.
Original record; inspected access route.
Actual PMC main Methods 9–17 and Results/Discussion18–46, main Figure1–4 captions read. Endogenous human HEK293 PTG–laforin coIP is separate from CHO-IR overexpression, substrate-level glycogen measurements and biochemical reconstitution. Figure 3C malin-dependent PTG ubiquitination requires recombinant laforin and fails with malin E280K; removal of laforin abolishes the reaction. This supports adaptor activity without attributing E3 chemistry to laforin. C266S retains cellular PTG effects. One sentence about direct PTG–malin binding conflicts with the preceding failed direct-binding assays; the assay-specific account is retained. No figure pixels/supplement.
Original record; inspected access route.
Normal/PubMed abstract read; source describes cytoplasmic/nuclear laforin isoforms and anL310W effect limited to the cytoplasmic product. Complete original Methods/Results unavailable after bounded access. Broad cytoplasm is independently supported; the membrane-specific assertion remains unresolved. Neither host species nor exact product sequence is inferred.
Original record; inspected access route.
Normal/PubMed abstract read, with explicit distinction of331/317-aa isoforms and reported differential phosphatase/glycogen-binding behavior. ROOT independently found OUP article and ostensiblyOA PDF redirect to the purchase/abstract page. Operative assays were not read, so no blanket monomer/dimer requirement or functional equivalence is imported. Positive later human 331-aa solution studies support dimerization under their own conditions.
Original record; inspected access route.
Actual PMC XML Methods 4–23 and operativeResults 37–57 plus main captions read; clinical descriptions were inspected only as context. Human malin identity is explicit, laforin constructs reference prior work. Two-hybrid interaction with expression controls and HEK293 PTG/glycogen effects support the malin-laforin complex. COS7 localization does not establish a simian laforin construct. No pixel/supplement inspection.
Original record; inspected access route.
Normal/PubMed abstract read and bounded original access independently repeated by ROOT; only request-full-text and citation pages recovered. It reports three transcripts/five possible translated proteins, differing localization and interactions, but the exact original constructs/reading-frame assays remain unread. All eight imported products, including external344-aaB3EWF7-1, are retained separately. No functional equivalence or exact per-product assay mapping is guessed.
Original record; inspected access route.
Actual PMC Methods 12–24, Results 59–67, Figures 6–7 captions and Discussion 72–79 read. Tagged coexpression/coIP in COS7/HeLa supports a multicomponent association and PRDM8-dependent nuclear foci. EndogenousPRDM8 pulls down a prior human C266S myc-laforin transgene from mouse tissues; not endogenous human WT binary binding. Source reports a complex, not a specific transcriptional activity of laforin. No supplementvideo or image pixels read.
Original record; inspected access route.
All40 actual PMC main indexed blocks read, including Methods 11–16, Results17–29 and Discussion31–33. Human 331-aa full-length preparation is distinct from DSP150–331C266S crystal and CBM1–137 constructs. AmylopectinPi assay is distinct from OMFP. AUC/SAXS supports solution dimer; C329S remains dimeric, so one disulfide mechanism is not universalized. No pixel inspection.
Original record; inspected access route.
Independent ROOT sealed actual PMC Methods/Results/Figures 1–7 and Discussion read scope. Figure 1C measuresPi release from rabbit-muscle glycogen by WT versus C266S/VHR. Main cloning:human 1–328N-His6; DXMS separately describesC-His6. Solution measurements supportdimer. Maltohexaose orientation predictsC2/C3 proximity rather than directly measuringC2-specific catalysis. Linked supplement returnedHTML and was not read; no pixels.
Original record; inspected access route.
Actual PMC STARMethods147–179 read: human ORFeome/OC baits, C-terminalFLAG-HA, separate293T/HCT116 experiments, AP-MS quality controls and gene-level aggregation. The exactEPM2A–NHLRC1 supplementary record/clone was not independently extracted, so isoform identity or binary mechanism is not inferred from this source. Independent15930137 directly supports malin association. This is a partialmain read, not absent full-text access.
Original record; inspected access route.
Normal abstract identifies a human disease gene encoding a predicted protein tyrosine phosphatase. This is not treated as an independently inspected glycogen-catalysis experiment. Direct later25544560 phosphate-release measurements establish glycogen metabolic participation. No exhaustive clinical/sequence-source original sweep was attempted.
O95278 and the separate reviewedB3EWF7 record remain distinct. B3EWF7-1 is 344 aa; it is not manufactured by truncating O95278. Its four raw GOA assertions remain outside the 81-row O95278 seed. The acquired source records are preserved even where source-to-product mapping remains unresolved.
PMID:20453062(https://doi.org/10.1093/hmg/ddq190) actual Methods/Results and main captions were read (XMLblocks7–51). Human patient fibroblast and Epm2a-null mouse/MEF flux assays, together with laforin expression and TSC2-null epistasis, support a gene-level autophagy/mTOR regulatory context. They do not identify a 344-aa POCR-specific mechanism. The Myc-laforin construct cites a previous Cancer Cell paper; the previous construct original was not recovered. Do not transfer these results indiscriminately to all products or propose a new canonical process solely because an external record carries it.
PMID:24837458(https://doi.org/10.1042/BSR20140053) actual Methods10–21, Results39–42/Figure4 andDiscussion61 were read. Human HepG2-derived STBD1 is distinct from the FLAG-laforin construct lineage through17908927/15930137 to11739371 human 331-aa. COS-cell co-IP and endogenous mouse-liver GST-STBD1 pull-down support association but do not establish a 344-aa POCR assay or invariant binary binding; glycogen bridging remains possible. This is a source/product-mapping question, not a title-only wrong-gene assertion. No supplemental data/pixels were inspected.
This assessment used actual official rule, donor, reaction, physical-entity and localization records, then compared their claims with the original experiments listed below. The normal publication cache is a separate immutable acquisition record; its access flag does not describe independently recovered originals. No new publication quotations, image-pixel readings or supplementary-material readings were added in this lane.
| Original | Actual operative scope and biological boundary |
|---|---|
| PMID18852261, DOI, PMC original | Methods glycogen-synthase assay; Results/Figures 4 and 6. Mouse Epm2a loss changes glycogen and GYS measurements. The glucose-transfer assay measures glycogen synthase in extracts; laforin removes phosphate. |
| PMID18040046, DOI, PMC original | Methods protein preparation; Results/Figures 1–4. Recombinant mouse laforin acts on glycogen/amylopectin. Separate mouse knockout measurements do not establish human construct identity. |
| PMID22669944, DOI, PMC original | Methods fractionation/immunogold; Results/Figures 4–6. Native mouse free and glycogen-associated laforin; recombinant mouse C265S in HEK293 is still mouse protein. |
| PMID17118331, DOI, PMC original | Methods and Results on C266S transgenic mice and GYS/branching-enzyme measurements. High overexpression and tissue-specific effects limit interpretation; GYS activity is not laforin catalysis. |
| PMID28536304, DOI, PMC original | Results/Figures 1–5 and phosphate/chain-length Methods. Human C266S and mouse C265S transgenes are distinguished. Phosphatase-independent rescue of chain abnormalities/Lafora bodies depends on malin and occurs despite persistent glycogen hyperphosphate. |
| PMID31042462, DOI, PMC original | Mouse muscle fractionation, Results/Figure 4, glycogen/GYS Methods and study limitations. Soluble GYS activity and insoluble accumulation are separate readouts. Human GBE1 expression is not a human laforin assay. |
| PMID11001928, DOI, author-upload access | Results p2252/Figure 2B; Methods pp2258–2259: human 331-aa GST-laforin releases phosphate from separately Ser/Thr- or Tyr-labeled myelin basic protein. This eight-hour protein assay differs from OMFP. Localization Results/Figures 6–8 support polysome association, not demonstrated membrane attachment. |
| PMID11739371, DOI, author-upload access | Experimental Procedures and Results/Figure 2B–D. Human muscle-derived laforin, protein-free glycogen cosedimentation and cellular fractionation were distinguished. pNPP is a separate synthetic substrate. |
| PMID15102711, DOI, author-upload access | Transgene-generation Results; brain/liver Results and Figures 1–7 captions. Human myc-C266S overexpression in mice supports soma/dendrite signal. Near-ER signal may be polyglucosan-associated. Native human biopsy and recombinant starch/glycogen binding are separate experiments. |
| PMID25538239, DOI | Actual PMC original Methods/Results, indexed blocks 11–29: human full-length 331-aa laforin, direct amylopectin phosphate release, AUC/SAXS dimerization. The inactive isolated DSP150–331 crystal and separate OMFP assay are not full-length glucan experiments. |
| PMID16901901, DOI | Actual PMC original Methods and Results: amylopectin versus pNPP, negative GSK3/PTG/protein-panel tests and CBM-fusion controls. Negative tested substrates do not exclude all possible protein substrates. |
| PMID17908927, DOI | Actual PMC original Methods and Figure 3C: tagged laforin in HepG2 cells partially enters the nucleus under glycogenolytic conditions. The separately described mouse AGL construct is not laforin. |
| PMID22961547, DOI | Actual PMC original Methods and Figures 6–7: tagged laforin localization and PRDM8-dependent nuclear redistribution in HeLa/COS7. Does not establish constitutive localization for every product. |
| PMID18070875, DOI | Actual PMC original Methods and Figures 1–3: CHO-IR glycogen readouts, endogenous HEK293 PTG–laforin complex and laforin-dependent reconstituted PTG ubiquitination are separate. Malin is the ligase; laforin contributes a scaffold. |
PMIDs16971387,15541350 and24430976 were identifier/abstract verified but their original operative texts were not recovered in this lane. Their donor records were not erased or treated as false solely because access failed; independent inspected originals support relevant target chemistry where possible. PMIDs11220751/11883934 remain an owner-reported original-access gap relevant to plasma-membrane assertions.
Human Protein Atlas EPM2A reports supported nucleoplasm localization with HPA053643/HPA055468 in SH-SY5Y/U2OS, and no staining for HPA055468 in SK-MEL-30. Actual XML/page annotations were read; no image pixels or product-specific assignment were inferred.
The mouse Q9WUA5 record identifies Epm2a and ENSMUSP00000066050.5. Current QuickGO donor assertions were traced individually. This lane contains no IBA rows and makes no PAINT-node assertion. Eleven ARBA rules had actual matching target conditions; InterPro and Rhea records were inspected separately from experimental substrate proof.
For Reactome R-HSA-3781011, catalyst3781002 identifies activeUnit3777103, whose referenceEntity146927 is human O95278. The same target and its cytosol compartment are traceable through the other assigned binding, dephosphorylation and ubiquitination events. GYG2 events inferred from GYG1 are labeled as such; FailedReaction objects do not establish mutant activity. R-HSA-3322077 contains both chain-building and associated maintenance events, so pathway membership does not confer GYS glucose-transfer catalysis on laforin.
Current GO:0019203 remains appropriate for carbohydrate phosphatase; inspected children provide no verified polymer-specific alternative. GO:0016311 has the current usage restriction None, which means it should not be used for direct annotation; this is not an annotation count. The term is non-obsolete but marked as an obsoletion candidate. No suitable glucan child was established, so no replacement was authored. GO:2001069 is the verified glycogen-binding term.
The external B3EWF7-1/POCR four GOA records remain separate from the 81 O95278 source rows. Product-specific attribution requires owner inspection of construct lineages; canonical 331-aa assays and whole-locus knockout phenotypes are not automatically POCR assays. No source rows or product definitions were changed.
Official PMC identity verified. Full main Experimental Procedures, Results/Figures 1-6 and Discussion were read. Recombinant full-length human laforin and W32G are distinguished from Arabidopsis SEX4/LSF2 variants. Selectively C3- or C6-radiolabeled Arabidopsis starch establishes C3 preference with activity at both positions, not exclusivity or measured C2 specificity. Soluble potato amylopectin/phospho-oligosaccharides and insoluble starch assays demonstrate substrate-dependent effects of the carbohydrate-binding domain. Chemical phosphate release is direct catalytic evidence; no clinical or all-isoform equivalence is inferred. No figure pixels or separate supplement were inspected. PMID:26231210(https://pubmed.ncbi.nlm.nih.gov/26231210/). The glycogen substrate assay supporting direct glycogen metabolism is PMID:25544560 Figure 1C, with separate inactive-C266S and VHR controls. The adaptor unit is supported by PMID:18070875 Methods and Figure 3C: laforin presence/absence and interaction-impaired malin E280K distinguish coordinated substrate ubiquitination from association alone. No additional process annotation is proposed.
GO:1990756 currently has the official label ubiquitin-like ligase-substrate adaptor activity and is an unrestricted molecular function. Its definition includes ubiquitin ligases, and its is_a path reaches GO:0030674 through GO:0140767. The specific term fits PMID:18070875 Figure 3C, with laforin addition/removal and malin E280K controls, and replaces the more general adaptor term in the PTG rows and second core. This does not imply UFM1 chemistry for laforin. PMID:22961547 Methods and Figures 6–7 support the malin partner-class binding refinement within the PRDM8-containing complex, without converting a co-immunoprecipitation into a purified binary contact.
The official QuickGO display template, lines 117–122 in the retrieved source, explicitly maps usage None to a restriction against direct annotation. The GO decision 26424 and current OLS term record corroborate the dephosphorylation restriction. Existing imported term identifiers are preserved.
The protein-process follow-up reread PMID:11001928 Methods pp. 2258–2259 and Figure 2B, PMID:11739371 Experimental Procedures/Results/Figure 2, and PMID:16901901 Methods/Results/Figures 1–4. The positive eight-hour MBP chemistry is retained as molecular capacity; the process over-annotation judgment is limited to the unsupported physiological extension. The HIRIP5 original remains unavailable and is not refuted by negative results on other substrates. No additional figure pixels or supplements were inspected in this follow-up.