The genuine Seed8 input contains 18 source annotations, including the explicit NOT annotation for calcium-gated chloride-channel activity. The original review and raw files were preserved before authorship in tmp/ANO5-initial/baseline/. Existing publication, Reactome and family caches are read-only. This journal records manual research; it is not provider-generated deep research.
One configured Falcon attempt with the installed client failed authentication; the configured Perplexity fallback failed DNS. Neither produced a report. A single normal seven-PMID cache batch into a temporary directory then timed out after 45 seconds without output. This is one interrupted batch, not seven completed individual failures. Receipts and logs are in tmp/ANO5-initial/.
The full cached PMID:33496727, ANO5 ensures trafficking of annexins in wounded myofibers, was read in the targeted Results, figure legends, Discussion and cDNA/imaging Methods. These are mouse isolated fibers with human ANO5 rescue constructs, plus myocytes from a human R58W patient; a human culture host alone is not being used to establish construct species. The cDNA Methods explicitly identify WT human ANO5 and the human ANO5/mouse ANO1 scrambling-domain chimera. WT human ANO5 restored repair to Ano5-deficient fibers, localized at wound-adjacent membrane and supported the normal annexin response. Human mutant myocytes had impaired repair.
The source directly separates repair from scrambling: "Both ANO5 and ANO5-1-5 expression in ANO5-KO myofibers restored normal repair" PMID:33496727. The chimera lacks the scrambling domain. It rescued ANXA6 trafficking and repair without restoring normal ANXA2 shedding. Lipid exposure in wounded adult fibers persisted without ANO5 and in calcium-free conditions. Thus repair is an established ANO5 role, but it must not be drawn as a process directly executed by its scramblase activity on this evidence. Direct ANO5–annexin binding, membrane-bending chemistry, and vesicle-fusion catalysis were not established. The authors acknowledge that dye entry alone cannot distinguish membrane fragility from delayed repair and discuss the remaining mechanism limits.
The canonical PMID:15124103, The novel gene encoding a putative transmembrane protein is mutated in gnathodiaphyseal dysplasia (GDD), is abstract-only. Its abstract explicitly reports ER localization and human GDD variants. Its proposed calcium-homeostasis explanation is a hypothesis, not a direct calcium-channel assay.
PMID:20056604, Expression and function of epithelial anoctamins, is also abstract-only in the canonical cache. The abstract reports plasma-membrane localization of all ten overexpressed anoctamins in rat thyroid cells, while ANO5 was not among proteins producing a positive calcium-activated halide-quenching result. This supports retaining the experimental localization without equating location with channel activity.
PMID:22075693, ANOs 3-7 in the anoctamin/Tmem16 Cl- channel family are intracellular proteins, has an abstract-only cache. The abstract reports no calcium-activated chloride currents from ANO3–7 in HEK293 cells. Targeted indexed original Results on ANO1/ANO5 chimeras were additionally read at the primary PMC record; those constructs failed surface trafficking and current generation. This does not establish absence of all ANO5 ion permeability in every membrane or assay. Full original Methods were not recovered through that browser route.
PMID:22946059, Anoctamins are a family of Ca2+-activated Cl- channels, has an abstract-only cache. It reports transient currents after overexpression of ANO4–10, including cation permeability. The publisher full-text route did not open. Its positive ANO5 channel/transport assertions cannot be removed simply because another paper reports negative assays, or because later work calls ANO5 a scramblase. Source-specific selectivity and localization details remain to be reconciled.
The official GO:0005254 definition describes facilitated chloride movement through a pore; it does not require exclusive chloride selectivity. Nonselective permeability therefore does not by itself refute this term. Likewise, nonselective currents and scramblase activity can coexist.
10.1007/s00018-017-2704-9. The original publisher Results on localization, currents and selectivity, and construct/patch-clamp/scrambling Methods were read. Human 898-aa and 913-aa constructs differ in membrane expression. Human 898-aa ANO5 supports calcium-dependent phospholipid scrambling; both constructs produce nonselective currents under strong depolarization. These are cell assays, not purified reconstitution. A GDD variant increases activity. Do not invent an exact UniProt isoform suffix for the 898-aa construct.10.1085/jgp.201812097. PubMed abstract/figure-caption evidence reports scrambling, associated nonselective currents, deficient mouse precursor fusion and rescue. Full original body remains to be read after normal recovery. Fusion necessity alone will not manufacture a new fusion-process assertion.10.1038/s41420-019-0197-z. PubMed abstract and figure captions were read. Patient myoblasts carrying R758C have defective repair and calcium clearance; WT expression or buffering restores repair. ER-localized ANO5 and a homology model are shown. A modeled dimer is not an experimental ANO5 oligomerization measurement, and ER calcium clearance does not establish that ANO5 itself transports calcium.10.1172/JCI163808. Primary publisher Results/Figures 1–2 were read. Human endothelial siRNA experiments support ANO5-dependent surface phosphatidylserine exposure and procoagulant function, with controls for tissue-factor expression, cell death and calcium responses. This is endothelial evidence; platelet ANO6 biology must not be transferred to ANO5. The gene is not a coagulation protease.10.1038/ejhg.2012.224. PubMed abstract and Figure 4 caption were read: an independent negative ANO5 halide-transport assay accompanies human GDD genetics. The positive ANO1/ANO2 controls do not establish a universal ANO5 prohibition.10.1002/humu.24006. Citation and indexed abstract were checked; full text has not been read. This is a candidate source for assay/variant distinctions, not justification for a new bone-development process.10.1016/j.bbrc.2005.03.226. Citation identity and accessible beginning of the mouse cDNA abstract were checked. The detailed localization experiment still requires reading. It is a candidate for tracing mouse donor Q75UR0, not an already-verified exact Ensembl source chain.The normal PTHR12308 entries include ANO5 Q75V66 and ANO10 Q9NW15. This establishes family membership, not equivalent activities. No local exact PAINT IBD assertions for PTN000267334 or PTN001389043 were found; the seeded nodes and all donors remain intact, and ANO5's self donor is legitimate. The InterPro dimerization-domain label is present in the preserved UniProt record, but the live InterPro API route did not open; a label is not an independently read experiment.
The official GO:0017128 definition and parents describe ATP-independent transbilayer phospholipid movement, under intramembrane lipid carrier activity. A proposed molecular-function assertion would be grounded in the actual ANO5 assays, not family nomenclature. No new BP is proposed from disease severity, fusion necessity or a desire to fill a pathway. The existing broad phospholipid-translocation IBA can retain its appropriate scope. gocams/index.tsv has no Q75V66/ANO5 hit.
The 2024 primary study PMID:39337485(https://pubmed.ncbi.nlm.nih.gov/39337485/), DOI 10.3390/ijms25189998, was read in indexed Results 2.2, Discussion and Conclusions. It compares parental and ANO6-knockout HEK293 cells. ANO5 scrambling persists without ANO6, whereas currents and membrane expression vary with the background. Negative plasma-membrane scrambling for other paralogs does not exclude intracellular scrambling. The ANO10 owner will coordinate a single normal request for this shared source.
The original PMID:22946059 publisher article subsequently opened through its correct primary link. The earlier access statement above is superseded for this source. Relevant Results, Figures 3–5 and 8 legends, Discussion, and construct/immunostaining Methods were read; all supplementary images were not inspected. The Methods explicitly identify human TMEM16E and the other human constructs. Ionomycin evokes small transient currents after ANO5 expression, and surface staining corroborates the experimental plasma-membrane assertion. The measured anion-ranking subset comprises ANO1, ANO4, ANO6 and ANO10, not ANO5. Endogenous ANO6 background and possible accompanying cation conductance are acknowledged. Neither an ANO5-specific selectivity ranking nor a purified ANO5 channel experiment is being inferred.
Accordingly, the prospective second assessment retains the positive channel and ion-transport assertions as secondary, assay-dependent functions. The negative source assertion from PMID:22075693 remains unresolved with its NOT flag intact: negative current under a particular expression/localization condition need not exclude activity under another. The exact ARBA rule remains untraced, although human target experiments independently support the broad channel assertion. The distinct scramblase function remains the proposed molecular-function core. This refines the original proposal rather than treating later source access as though it had been available throughout.
The official indexed mouse UniProt Q75UR0 entry additionally resolves the donor as mouse Ano5, links ENSMUSP00000046884.6, and lists an MGI-derived vesicle annotation. This establishes donor identity and the existence of the source annotation, not its original localization experiment or the full Ensembl transfer event. The vesicle decision remains unresolved pending that distinction.
The coordinated ANO10 normal request also includes PMID:32517157(https://pubmed.ncbi.nlm.nih.gov/32517157/), Targeting of Intracellular TMEM16 Proteins to the Plasma Membrane and Activation by Purinergic Signaling, DOI 10.3390/ijms21114065. Its official abstract was read: ANO5 and other intracellular paralogs are artificially targeted with a CAAX motif. Engineered surface localization cannot establish native distribution, and absent extra surface scrambling in those constructs does not by itself refute the other ANO5 assays. No second normal fetch was made for either shared source.
All nine relevant normal records (seven ANO5 requests and two shared records) were read directly from the exact Source32 archive before import and then verified byte-identical to the root-imported canonical records. Eight of eleven batch records have XML full-text flags; for this gene six of nine do. PMID:15882990, PMID:23047743 and PMID:32112655 remain abstract-only. In particular, PMID:23047743 XML was restricted and its PDF fallback yielded only 54 characters. Flags were preserved, and actual source scopes—not flags alone—govern the review.
The earlier inaccessible-Methods statements are superseded for PMID:22075693: the original primary article identifies human ANO5 IMAGE100061756 with C-terminal EGFP and negative whole-cell currents at about 24µM calcium. Its Discussion itself leaves intrinsic CaCC function unresolved owing to intracellular retention. The NOT row remains UNDECIDED for this scientific context, not because those Methods remain unread. The original PMID:20056604 cDNA Methods explicitly identify human ANO5 from A549 cells; rat FRT is the host. Targeted His/β-catenin localization and endogenous-background Results were read; no complete supplement read is claimed. Both canonical caches remain abstract-only.
All 18 original source objects, the NOT qualifier and absent alternative-product field remain intact. Decisions remain 10 ACCEPT, 5 KEEP_AS_NON_CORE, 3 UNDECIDED, plus one NEW scramblase MF and one core. No new BP, isoform suffix, intrinsic calcium-pump claim or repair-from-scrambling causal link was added. The vesicle donor chain and dimerization mapping remain unresolved; the actual PMID:31341644 dimer is explicitly a homology model. Source PMID:29124309 current measurements remain secondary/conditional, with endogenous-background caveats from PMID:39337485. Normal cache closure adds exact literal citations and source-specific read limits; it does not make every historical annotation certain.
The published baseline is commit 0b0ddad176cf6e937ea358a9be211fab3904f9b7. All 24 scoped files were compared byte-for-byte with that commit before editing. The 18 original GOA source objects, including the NOT flag, and all 19 decisions are preserved. The existing NEW scramblase row now explicitly carries enables; no additional annotation or process was proposed. The parent independently assessed the channel and repair evidence before these changes (tmp/ANO5-root-followup-prospective.json).
The current official definitions of GO:0005254 and GO:0005229 require chloride passage, with calcium-responsive gating for the latter; they do not require exclusive chloride selectivity. The positive source-specific rows remain NON_CORE after weighing the contrary studies explicitly. This does not establish a native ANO1-like epithelial channel function. The ARBA rule itself remains unresolved; retaining its broad term rests on independent human target experiments, not a newly reconstructed rule.
I reread the targeted current/selectivity/calcium Results and Figures 2–4 legends and construct Methods of PMID:29124309(https://pubmed.ncbi.nlm.nih.gov/29124309/). Human 898- and 913-residue constructs produced currents in HEK cells, and the shorter construct was also tested in CHO cells. Large currents at 3 µM intracellular calcium required strong positive potentials; the full-length construct targeted the surface less efficiently. Matched controls did not develop comparable currents. Sodium/chloride substitution and reversal-potential experiments support poor selectivity, and calcium favors activation without being an absolute requirement. These results support conditional activity without resolving its native physiological contribution.
The negative FRT halide assay in PMID:20056604(https://pubmed.ncbi.nlm.nih.gov/20056604/), intracellular retention and negative whole-cell assay in PMID:22075693(https://pubmed.ncbi.nlm.nih.gov/22075693/), and contrary channel interpretation in PMID:23047743(https://pubmed.ncbi.nlm.nih.gov/23047743/) now appear directly in the channel reasoning. Complete cached abstracts were reread; prior external article access remains limited to the sections recorded above. The original negative result is preserved. Its broader NOT interpretation remains UNDECIDED because construct, localization and stimulation conditions differ across the studies. No unread negative source was dismissed.
Actual PMID:39337485(https://pubmed.ncbi.nlm.nih.gov/39337485/) Results 2.1–2.3 and Discussion were reread. Its Discussion links ANO5/ANO10 currents to ANO6 absence, while ANO9 currents depend on ANO6 presence. This is evidence of expression-background dependence, not proof that all ANO5-associated currents are carried by endogenous ANO6. The current conclusion refers to Figure S1C in the authors' text; I did not newly inspect those supplementary panels. ANO5-associated lipid exposure remains measurable without ANO6, with a smaller overall signal. Construct species is still not inferred from the human host.
Reusable findings now record the mutation-direction result from the complete abstract of PMID:32112655(https://pubmed.ncbi.nlm.nih.gov/32112655/) and the engineered-targeting limit in PMID:32517157(https://pubmed.ncbi.nlm.nih.gov/32517157/). For the latter, I reread targeted Results 2.1–2.3, Discussion and cell/construct Methods: CFP-CAAX forced surface localization supports current/anion-permeability responses but no added scrambling in the tested conditions. That is a relevant negative finding for these engineered constructs, not a universal negative for non-CAAX ANO5. The positive human ANO5 cell assays and endothelial knockdown evidence still support the existing scramblase MF.
Membrane repair remains ACCEPT and biologically central. PMID:33496727(https://pubmed.ncbi.nlm.nih.gov/33496727/) directly shows repair rescue by both ANO5 and ANO5-1-5, separating repair from the scrambling domain. Targeted repair/annexin Results and the Discussion also leave a mechanistic boundary: localization and annexin redistribution do not identify a direct ANO5 binding, scaffold, transport or fusion activity that executes repair. The core description now states this reason for omitting a separate repair MF, and a literature-grounded knowledge gap captures the unknown step. The schema has no proposed_molecular_function slot; no unsupported replacement activity was invented. Plasma membrane in the scrambling core identifies where surface lipid exposure was measured and does not erase the established ER pool.
Reference-review prose was shortened to biological findings and actual access limits. Fifteen exact, source-specific findings were added across the relevant publications. Disease names and anoctamin-family membership in the description are supported by the preserved UniProt record. All cached titles, publication bytes, raw GOA/UniProt and earlier histories remain unchanged.