The genuine normal intake contains 33 source assertions, two alternative products and 15 reference identities. All are retained exactly. The provider order is unusual but authentic: product name 1 is the displayed 816-residue P51795-2, and product name 2 is P51795-1, with the first 70 canonical residues removed. Historical 746-residue constructs are not assigned to a current product merely from their length. The original GOA, UniProt and provider artifacts remain immutable.
The normal Falcon attempt timed out after 90 seconds; the actual Perplexity fallback returned an insufficient-quota error. Manual research therefore used authentic primary sources and official annotation services. Existing main caches were retained byte for byte. Five shared paths were authenticated against the already published CLCN4 review; the genuine alternative timestamp-bearing family metadata remains archived. The accompanying source index contains identifiers, URLs, hashes, relevant literal interaction fields and assay scopes. It is provenance, not independent experimental support.
The early human study links CLCN5 variants with renal stone disorders and reduced oocyte currents. Its complete abstract was read, but the original full article was not recovered; a repository route also returned an access challenge. Those observations are preserved as positive current and renal-function evidence, not represented as proton-flux measurements PMID:8559248. Later original ClC-4/ClC-5 experiments directly measure proton transport and distinguish coupled exchange using gradient, channel and uncoupling controls. Their stoichiometric estimate is approximate, not a precise universal 2:1 measurement PMID:16034422. This supports refining the historical channel, generic transporter and antiporter labels to chloride:proton antiporter activity (GO:0062158), without rewriting the source assertions.
Mouse E211A knock-in experiments separate exchange from chloride conduction: uptake defects persist despite preserved bulk ATP-driven vesicular acidification. The original article and selected supplementary Methods include littermate, localization and uptake controls. The chloride-accumulation interpretation is a model rather than a directly measured endosomal chloride result in that study PMID:20430975. A separate human-expression study measures a bafilomycin-insensitive acidification component in HEK293 vesicles, with transport mutants as controls; its opossum-cell knockdown and proton-entry interpretation remain distinct PMID:20421284. Human ClC-5 oocyte measurements provide additional direct transport evidence, with apparent stoichiometry calculated from electrochemical energetics and epithelial localization evaluated separately PMID:31852738. These context-dependent findings motivate a question about native voltage and ion gradients, not a categorical claim that ClC-5 always acidifies or always alkalinizes an endosome.
The existing renal process is retained because ClC-5 performs the ion-transport step supporting renal epithelial function. No new endocytosis, developmental or disease process is added solely from loss-of-function and rescue phenotypes. The single organizing core is endosomal chloride/proton exchange with the established renal role.
The recovered original renal study examines human fetal and postnatal kidneys alongside mouse development. Human fetal proximal-tubule staining is less polarized; detailed apical/subapical comparisons include mouse sections. These species and developmental distinctions are retained in the apical annotation PMID:14675051. Human Protein Atlas records independently support Golgi and plasma membrane pools in CACO-2 and RH-30 cells, with plasma membrane/cytosol in U-2 OS. Selected images were inspected, but no quantitative colocalization analysis was performed. The surface pool is experimentally real and varies with trafficking context; it is not made the principal physiological compartment.
The exact human ClC-5–KIF3B evidence includes two-hybrid, pull-down and reciprocal co-immunoprecipitation assays. Six literal interaction records contain five association assays and one confocal colocalization record. The latter is not counted as a binding assay. Paper tail numbering 551–746 and curated record numbering 621–816 are both retained. The GST pull-down uses a bacterial partner preparation and an in-vitro-translated tail; it is not described as two purified full-length proteins. Separate endogenous mouse-kidney work supports a kinesin-2 association. KIF3B-dependent surface and uptake effects differ between HEK293 and polarized opossum kidney cells. Kinesin binding is therefore a specific supported refinement, without attributing motor or ATPase activity to ClC-5 PMID:19940036. Newer TMEM9-family experiments retain their human-transporter, mouse-partner, renal-tissue and engineered-tag boundaries PMID:40169677.
The original lysosomal proteomics result is positive: official Table 1 identifies human placental ClC5, IPI00294056, with seven peptides, 12.9% sequence coverage and 31 spectra. Its logarithmic enrichment statistic is 2.44 and adjusted P value is 1. Detection is not equivalent to statistically significant enrichment, and fluorescence validation addressed other proteins. Peptide supplement PDFs were inaccessible, so no isoform-discriminating detection is claimed. This source is retained as non-core PMID:17897319. The distinct NK-cell HDA assertion remains UNDECIDED because its exact target table and peptides were not recovered. An abstract describing the general proteome cannot resolve that source-specific observation PMID:19946888.
The original human cytoplasmic-domain study directly measures binding of three adenyl ribonucleotides by equilibrium dialysis and resolves ATP- and ADP-bound structures. Binding-site mutants provide further specificity controls. Its Methods gives residues 568–746, whereas the Results gives 571–746; no exact modern product mapping is inferred. One NEW GO:0032559 annotation captures the shared ligand class without nested ATP/ADP/AMP annotations PMID:17195847.
Sedimentation supports weak self-association of the isolated domain, unchanged by 5 mM ATP, while the authors leave native domain geometry unresolved. The existing identical-protein-binding annotation is retained as non-core. Wild-type-background binding-site mutants have unchanged currents; effects in the uncoupled E211A background do not establish physiological nucleotide gating. The publisher PDF includes a correction exchanging the original Figure 4d labels, and the independent consultation used the corrected figure. No ATPase activity, universal native dimer arrangement or new gating process is proposed.
PAINT nodes are treated as curated phylogenetic assertions, not pairwise similarity transfers. Current donor and target experimental records provide corroboration; the historical tree, MSA and IBD placement were not reconstructed. Target self-inclusion is valid experimental grounding, and donor count is not used as a confidence score. Original is_active_in qualifiers remain exact; staining and fractionation are not presented as direct flux assays in every compartment.
Current ARBA00026476 conditions match the target in two sets and emit an older channel term. The two other current rule snapshots do not reproduce the historical target route; this is explicitly distinguished from biological absence. Reactome resolves P51795, residues 1–816, as an alternative member of an endosomal exchange catalyst set. CLCN4, CLCN5 and CLCN6 are not an obligatory three-protein complex. Typed ontology coverage shows that the one proposed ligand-binding activity is neither an ancestor nor a descendant of a seeded assertion. The authenticated GO-CAM index has no target activity; this absence is not used to manufacture a process annotation.
The membrane annotation from PMID:14675051 is retained as ACCEPT at its original resolution. The study's human material includes a bulk membrane pellet and developmental segmental staining; an endosome-specific replacement cannot be assigned to that experiment merely from another source. The separate endosome annotations remain unchanged.
The earlier construct-mapping uncertainty above is superseded by sequence comparison. P51795-1 (746 residues) equals P51795-2 (816 residues) after removal of residues 1–70, including the E211→E281, Y617→Y687, S618→S688 and D727→D797 correspondences. The original Methods range 568–746 maps to displayed 638–816, while Results 571–746 maps to 641–816. Native residues in deposited 2J9L and 2JA3 match 641–816 exactly, with an N-terminal methionine and an eight-residue C-terminal expression extension. The PDB record labels the added residues as expression tags. UniProt's curated crystallography range is 640–816; these distinct boundaries are not explained as rounding. The domain is shared by both products, but the experiment does not separately assay their full-length physiological activity [PMID:17195847; PDB:2J9L; PDB:2JA3].
A bounded process audit supports one new endosomal lumen acidification assertion. Human ClC-5 expressed in HEK293 cells confers bafilomycin-insensitive vesicular acidification that localized proton-transport mutants do not; the measured performed step is proton translocation. Opossum-cell knockdown is corroborating context, not a human experiment PMID:20421284. The distinct mouse E211A experiment preserves bulk ATP-driven acidification despite defective renal uptake, so the addition does not make acidification the universal explanation of ClC-5-dependent renal absorption PMID:20430975. GO:0048388 is an active pH-reduction process with no typed overlap with the existing terms. Same-role CLCN3 has TAS support; CLCN4, CLCN6 and CLCN7 lack current annotations to this exact term. Thus it is not reserved to V-ATPases, but annotation absence is not the rationale for addition. The current GO-CAM index has no target hit. No broader pH process is added. The core uses the more specific renal absorption term for the established renal role.
TMEM9-family experiments show association, inhibited currents and altered trafficking of human ClC-5 in heterologous systems with mouse TMEM9 constructs; mouse renal data distinguish transporter stability in tissue PMID:40169677. These findings inform the transport core and a question about human renal regulation. A separate generic binding assertion would add little mechanistic information, and ClC-5 is not assigned the inhibitor activity performed by its partners.
The original notes remain above as a chronological record. Reference assessments now emphasize access and verification scope rather than repeating their findings. All incoming source assertions, product objects, reference identities, source caches and history remain preserved; the explicit changes are one corrected membrane decision, the supported process addition, and the stated synthesis refinements.