[1]	NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS 1 AND 2), FUNCTION, TISSUE SPECIFICITY, DEVELOPMENTAL STAGE, AND SUBCELLULAR LOCATION. TISSUE=Kidney; DOI=10.1038/36285; PubMed=9363890 [NCBI, ExPASy, EBI, Israel, Japan] Kuro-o M., Matsumura Y., Aizawa H., Kawaguchi H., Suga T., Utsugi T., Ohyama Y., Kurabayashi M., Kaname T., Kume E., Iwasaki H., Iida A., Shiraki-Iida T., Nishikawa S., Nagai R., Nabeshima Y.; "Mutation of the mouse klotho gene leads to a syndrome resembling ageing."; Nature 390:45-51(1997).
[2]	NUCLEOTIDE SEQUENCE [GENOMIC DNA], ALTERNATIVE SPLICING (ISOFORMS 1 AND 2), AND TISSUE SPECIFICITY. DOI=10.1016/S0014-5793(98)00127-6; PubMed=9537505 [NCBI, ExPASy, EBI, Israel, Japan] Shiraki-Iida T., Aizawa H., Matsumura Y., Sekine S., Iida A., Anazawa H., Nagai R., Kuro-o M., Nabeshima Y.; "Structure of the mouse klotho gene and its two transcripts encoding membrane and secreted protein."; FEBS Lett. 424:6-10(1998).
[3]	TISSUE SPECIFICITY, AND SUBCELLULAR LOCATION. DOI=10.1006/bbrc.1999.2009; PubMed=10631108 [NCBI, ExPASy, EBI, Israel, Japan] Kato Y., Arakawa E., Kinoshita S., Shirai A., Furuya A., Yamano K., Nakamura K., Iida A., Anazawa H., Koh N., Iwano A., Imura A., Fujimori T., Kuro-o M., Hanai N., Takeshige K., Nabeshima Y.; "Establishment of the anti-Klotho monoclonal antibodies and detection of Klotho protein in kidneys."; Biochem. Biophys. Res. Commun. 267:597-602(2000).
[4]	FUNCTION. DOI=10.1006/bbrc.2000.3864; PubMed=11095966 [NCBI, ExPASy, EBI, Israel, Japan] Mori K., Yahata K., Mukoyama M., Suganami T., Makino H., Nagae T., Masuzaki H., Ogawa Y., Sugawara A., Nabeshima Y., Nakao K.; "Disruption of klotho gene causes an abnormal energy homeostasis in mice."; Biochem. Biophys. Res. Commun. 278:665-670(2000).
[5]	FUNCTION. DOI=10.1053/meta.2000.8606; PubMed=11016890 [NCBI, ExPASy, EBI, Israel, Japan] Utsugi T., Ohno T., Ohyama Y., Uchiyama T., Saito Y., Matsumura Y., Aizawa H., Itoh H., Kurabayashi M., Kawazu S., Tomono S., Oka Y., Suga T., Kuro-o M., Nabeshima Y., Nagai R.; "Decreased insulin production and increased insulin sensitivity in the klotho mutant mouse, a novel animal model for human aging."; Metabolism 49:1118-1123(2000).
[6]	INDUCTION, AND TISSUE SPECIFICITY. DOI=10.1016/S0024-3205(01)01092-X; PubMed=11411791 [NCBI, ExPASy, EBI, Israel, Japan] Mizuno I., Takahashi Y., Okimura Y., Kaji H., Chihara K.; "Upregulation of the klotho gene expression by thyroid hormone and during adipose differentiation in 3T3-L1 adipocytes."; Life Sci. 68:2917-2923(2001).
[7]	SUBCELLULAR LOCATION, AND TISSUE SPECIFICITY. DOI=10.1016/S0378-5955(02)00483-5; PubMed=12204354 [NCBI, ExPASy, EBI, Israel, Japan] Kamemori M., Ohyama Y., Kurabayashi M., Takahashi K., Nagai R., Furuya N.; "Expression of Klotho protein in the inner ear."; Hear. Res. 171:103-110(2002).
[8]	FUNCTION, AND INDUCTION. DOI=10.1210/me.2003-0048; PubMed=14528024 [NCBI, ExPASy, EBI, Israel, Japan] Tsujikawa H., Kurotaki Y., Fujimori T., Fukuda K., Nabeshima Y.; "Klotho, a gene related to a syndrome resembling human premature aging, functions in a negative regulatory circuit of vitamin D endocrine system."; Mol. Endocrinol. 17:2393-2403(2003).
[9]	INDUCTION. DOI=10.1016/j.cardiores.2004.07.011; PubMed=15485693 [NCBI, ExPASy, EBI, Israel, Japan] Narumiya H., Sasaki S., Kuwahara N., Irie H., Kusaba T., Kameyama H., Tamagaki K., Hatta T., Takeda K., Matsubara H.; "HMG-CoA reductase inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells."; Cardiovasc. Res. 64:331-336(2004).
[10]	SUBCELLULAR LOCATION, AND TISSUE SPECIFICITY. DOI=10.1247/csf.29.91; PubMed=15665504 [NCBI, ExPASy, EBI, Israel, Japan] Li S.-A., Watanabe M., Yamada H., Nagai A., Kinuta M., Takei K.; "Immunohistochemical localization of Klotho protein in brain, kidney, and reproductive organs of mice."; Cell Struct. Funct. 29:91-99(2004).
[11]	CLEAVAGE, TISSUE SPECIFICITY, SUBUNIT, GLYCOSYLATION, AND SUBCELLULAR LOCATION. DOI=10.1016/j.febslet.2004.03.090; PubMed=15135068 [NCBI, ExPASy, EBI, Israel, Japan] Imura A., Iwano A., Tohyama O., Tsuji Y., Nozaki K., Hashimoto N., Fujimori T., Nabeshima Y.; "Secreted Klotho protein in sera and CSF: implication for post-translational cleavage in release of Klotho protein from cell membrane."; FEBS Lett. 565:143-147(2004).
[12]	ENZYME ACTIVITY, ENZYME REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES. DOI=10.1074/jbc.M312392200; PubMed=14701853 [NCBI, ExPASy, EBI, Israel, Japan] Tohyama O., Imura A., Iwano A., Freund J.-N., Henrissat B., Fujimori T., Nabeshima Y.; "Klotho is a novel beta-glucuronidase capable of hydrolyzing steroid beta-glucuronides."; J. Biol. Chem. 279:9777-9784(2004).
[13]	FUNCTION, LACK OF ENZYMATIC ACTIVITY, TISSUE SPECIFICITY, AND SUBCELLULAR LOCATION. DOI=10.1126/science.1112766; PubMed=16123266 [NCBI, ExPASy, EBI, Israel, Japan] Kurosu H., Yamamoto M., Clark J.D., Pastor J.V., Nandi A., Gurnani P., McGuinness O.P., Chikuda H., Yamaguchi M., Kawaguchi H., Shimomura I., Takayama Y., Herz J., Kahn C.R., Rosenblatt K.P., Kuro-o M.; "Suppression of aging in mice by the hormone Klotho."; Science 309:1829-1833(2005).
[14]	PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT TYR-499, AND MASS SPECTROMETRY. TISSUE=Brain cortex; DOI=10.1074/mcp.M600046-MCP200; PubMed=17114649 [NCBI, ExPASy, EBI, Israel, Japan] Munton R.P., Tweedie-Cullen R., Livingstone-Zatchej M., Weinandy F., Waidelich M., Longo D., Gehrig P., Potthast F., Rutishauser D., Gerrits B., Panse C., Schlapbach R., Mansuy I.M.; "Qualitative and quantitative analyses of protein phosphorylation in naive and stimulated mouse synaptosomal preparations."; Mol. Cell. Proteomics 6:283-293(2007).

Comments

FUNCTION: May have weak glycosidase activity towards glucuronylated steroids. However, it lacks essential active site Glu residues at positions 241 and 874, suggesting it may be inactive as a glycosidase in vivo. May be involved in the regulation of calcium and phosphorus homeostasis by inhibiting the synthesis of active vitamin D.
FUNCTION: The Klotho peptide generated by cleavage of the membrane-bound isoform may be an anti-aging circulating hormone which would extend life span by inhibiting insulin/IGF1 signaling.
CATALYTIC ACTIVITY: A beta-D-glucuronoside + H₂O = D-glucuronate + an alcohol.
ENZYME REGULATION: Inhibited by D-saccharic acid 1,4-lactone and taurocholic acid.

BIOPHYSICOCHEMICAL PROPERTIES:

Kinetic parameters:	K_M=0.249 mM for 4-methylumbelliferylglucuronide;
	K_M=0.251 mM for estrone 3-beta-D-glucuronide;
	K_M=0.174 mM for beta-estradiol 3-beta-D-glucuronide;
	K_M=0.251 mM for estriol 3-beta-D-glucuronide;
	V_max=0.62 µM/h/ug enzyme;
pH dependence:	Optimum pH is 5.5;

SUBUNIT: Homodimer.
INTERACTION:
P04426:Wnt1; NbExp=1; IntAct=EBI-1570828, EBI-1570911;
P17553:Wnt3; NbExp=1; IntAct=EBI-1570828, EBI-1570853;
P22724:Wnt4; NbExp=1; IntAct=EBI-1570828, EBI-1570945;
P22725:Wnt5a; NbExp=1; IntAct=EBI-1570828, EBI-1570983;
SUBCELLULAR LOCATION: Isoform 1: Cell membrane; Single-pass type I membrane protein. Note=Isoform 1 shedding leads to a soluble peptide.
SUBCELLULAR LOCATION: Isoform 2: Secreted.

ALTERNATIVE PRODUCTS: 2 named isoforms [FASTA] produced by alternative splicing.

Name	1
Synonyms	Membrane-bound
Isoform ID	O35082-1
Note: Predominates over the secreted form by more than 10 times in all tissues examined.
This is the isoform sequence displayed in this entry.

Name	2
Synonyms	Secreted
Isoform ID	O35082-2
Features which should be applied to build the isoform sequence: VSP_015828, VSP_015829.

TISSUE SPECIFICITY: Membrane-bound protein is present in distal renal tubules, inner ear, ependymal cells of brain choroid plexus, elongating spermatids and mature oocytes (at protein level). Soluble peptide is present in serum (100 pM) and cerebrospinal fluid. Expressed strongly in kidney, moderately in brain choroid plexus, and at low levels in pituitary, placenta, skeletal muscle, urinary bladder, aorta, pancreas, testis, ovary, colon, thyroid gland and adipocytes.
DEVELOPMENTAL STAGE: Not expressed in the embryo. Expressed in the kidney of newborns.
INDUCTION: Induced by 1,25-dihydroxyvitamin D(3) in kidney. Down-regulated by angiotensin II and up-regulated by statins through modulation of the RhoA pathway in epithelial cells (in vitro). Isoform 1 (but not isoform 2) is up-regulated by thyroid hormone in adipocytes.
DOMAIN: Contains 2 glycosyl hydrolase 1 regions. However, the first region lacks the essential Glu active site residue at position 241, and the second one lacks the essential Glu active site residue at position 874.
PTM: N-glycosylated.
MISCELLANEOUS: Mice lacking Kl or with strong defects in Kl expression display a syndrome resembling to human aging, with short lifespan, infertility, arteriosclerosis, skin atrophy, osteoporosis and emphysema. They have various metabolic abnormalities, including increased insulin sensitivity and decreased insulin production. Mice overexpressing Kl have increased resistance to insulin and IGF1, a lifespan extended of more than 20%, and generate fewer offspring.
SIMILARITY: Belongs to the glycosyl hydrolase 1 family. Klotho subfamily [view classification].
WEB RESOURCE: Name=Protein Spotlight; Note=The thread of life -Issue 65 of December 2005; URL="http://www.expasy.org/spotlight/back_issues/sptlt065.shtml";.

Copyright

Copyrighted by the UniProt Consortium, see http://www.uniprot.org/terms. Distributed under the Creative Commons Attribution-NoDerivs License.

Cross-references

Sequence databases

EMBL

AB005141; BAA23381.1; -; mRNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
AB010088; BAA25307.1; -; mRNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
AB010091; BAA25308.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]
AB010091; BAA25309.1; -; Genomic_DNA.	[EMBL / GenBank / DDBJ] [CoDingSequence]

RefSeq

NP_038851.1; -.

UniGene

Mm.6500

3D structure databases

HSSP

Q08638; 1OIF. [HSSP ENTRY / PDB]

ModBase

O35082.

Protein-protein interaction databases

IntAct

O35082; -.

PTM databases

PhosphoSite

O35082; -.

Organism-specific databases

MGI

MGI:1101771; Kl.

Gene expression databases

ArrayExpress

O35082; -.

CleanEx

MM_KL; -.

GermOnline

ENSMUSG00000058488; Mus musculus.

Ontologies

GO:0005515;	Molecular function: protein binding (inferred from physical interaction from IntAct).
GO:0007568;	Biological process: aging (inferred from mutant phenotype from MGI).
GO:0006112;	Biological process: energy reserve metabolic process (inferred from mutant phenotype from MGI).
QuickGo view.

Family and domain databases

InterPro

IPR001360; Glyco_hydro_1.
IPR013781; Glyco_hydro_sub_cat.
Graphical view of domain structure.

Gene3D

G3DSA:3.20.20.80; Glyco_hydro_cat; 2.

PANTHER

PTHR10353; Glyco_hydro_1; 1.

Pfam

PF00232; Glyco_hydro_1; 3.
Pfam graphical view of domain structure.

PRINTS

PR00131; GLHYDRLASE1.

ProDom

PD000650; Glyco_hydro_euk; 2.
[Domain structure / List of seq. sharing at least 1 domain]

PROSITE

PS00572; GLYCOSYL_HYDROL_F1_1; FALSE_NEG.
PS00653; GLYCOSYL_HYDROL_F1_2; 1.

BLOCKS

O35082.

Genome annotation databases

Ensembl

ENSMUSG00000058488; Mus musculus. [Contig view]

GeneID

16591; -.

KEGG

mmu:16591; -.

Phylogenomic databases

HOGENOM

O35082; -.

HOVERGEN

O35082; -.

Other

Kl; Mus musculus.

View cluster of proteins with at least 50% / 90% / 100% identity.

Keywords

Alternative splicing; Cell membrane; Glycoprotein; Glycosidase; Hydrolase; Membrane; Phosphoprotein; Repeat; Secreted; Signal; Transmembrane.

Features

Feature table viewer

Feature aligner

`Key`	`From To`	`Length`	`Description`	`FTId`
`SIGNAL`	`1 34`	`34`	`Potential.`
`CHAIN`	`35 1014`	`980`	`Klotho.`	`PRO_0000042247`
`CHAIN`	`35 ?`		`Klotho peptide.`	`PRO_0000042248`
`TOPO_DOM`	`35 982`	`948`	`Extracellular (Potential).`
`TRANSMEM`	`983 1003`	`21`	`Potential.`
`TOPO_DOM`	`1004 1014`	`11`	`Cytoplasmic (Potential).`
`REGION`	`59 508`	`450`	`Glycosyl hydrolase-1 1.`
`REGION`	`517 955`	`439`	`Glycosyl hydrolase-1 2.`
`MOD_RES`	`499 499`		`Phosphotyrosine.`
`CARBOHYD`	`161 161`		`N-linked (GlcNAc...) (Potential).`
`CARBOHYD`	`285 285`		`N-linked (GlcNAc...) (Potential).`
`CARBOHYD`	`346 346`		`N-linked (GlcNAc...) (Potential).`
`CARBOHYD`	`609 609`		`N-linked (GlcNAc...) (Potential).`
`CARBOHYD`	`614 614`		`N-linked (GlcNAc...) (Potential).`
`CARBOHYD`	`696 696`		`N-linked (GlcNAc...) (Potential).`
`VAR_SEQ`	`537 550`		`DTTLSQFTDPNVYL -> SPLTKPSVGLLLPH (in isoform 2).`	`VSP_015828`
`VAR_SEQ`	`551 1014`		`Missing (in isoform 2).`	`VSP_015829`
`CONFLICT`	`854 854`		`L -> V (in Ref. 2; BAA25308).`

Sequence information

Length: 1014 AA [This is the length of the unprocessed precursor]

Molecular weight: 116426 Da [This is the MW of the unprocessed precursor]

CRC64: 24BD600D2F969FDC [This is a checksum on the sequence]

        10         20         30         40         50         60 
MLARAPPRRP PRLVLLRLLL LHLLLLALRA RCLSAEPGQG AQTWARFARA PAPEAAGLLH 

        70         80         90        100        110        120 
DTFPDGFLWA VGSAAYQTEG GWRQHGKGAS IWDTFTHHSG AAPSDSPIVV APSGAPSPPL 

       130        140        150        160        170        180 
SSTGDVASDS YNNVYRDTEG LRELGVTHYR FSISWARVLP NGTAGTPNRE GLRYYRRLLE 

       190        200        210        220        230        240 
RLRELGVQPV VTLYHWDLPQ RLQDTYGGWA NRALADHFRD YAELCFRHFG GQVKYWITID 

       250        260        270        280        290        300 
NPYVVAWHGY ATGRLAPGVR GSSRLGYLVA HNLLLAHAKV WHLYNTSFRP TQGGRVSIAL 

       310        320        330        340        350        360 
SSHWINPRRM TDYNIRECQK SLDFVLGWFA KPIFIDGDYP ESMKNNLSSL LPDFTESEKR 

       370        380        390        400        410        420 
LIRGTADFFA LSFGPTLSFQ LLDPNMKFRQ LESPNLRQLL SWIDLEYNHP PIFIVENGWF 

       430        440        450        460        470        480 
VSGTTKRDDA KYMYYLKKFI METLKAIRLD GVDVIGYTAW SLMDGFEWHR GYSIRRGLFY 

       490        500        510        520        530        540 
VDFLSQDKEL LPKSSALFYQ KLIEDNGFPP LPENQPLEGT FPCDFAWGVV DNYVQVDTTL 

       550        560        570        580        590        600 
SQFTDPNVYL WDVHHSKRLI KVDGVVAKKR KPYCVDFSAI RPQITLLREM RVTHFRFSLD 

       610        620        630        640        650        660 
WALILPLGNQ TQVNHTVLHF YRCMISELVH ANITPVVALW QPAAPHQGLP HALAKHGAWE 

       670        680        690        700        710        720 
NPHTALAFAD YANLCFKELG HWVNLWITMN EPNTRNMTYR AGHHLLRAHA LAWHLYDDKF 

       730        740        750        760        770        780 
RAAQKGKISI ALQADWIEPA CPFSQNDKEV AERVLEFDIG WLAEPIFGSG DYPRVMRDWL 

       790        800        810        820        830        840 
NQKNNFLLPY FTEDEKKLVR GSFDFLAVSH YTTILVDWEK EDPMKYNDYL EVQEMTDITW 

       850        860        870        880        890        900 
LNSPSQVAVV PWGLRKVLNW LRFKYGDLPM YVTANGIDDD PHAEQDSLRI YYIKNYVNEA 

       910        920        930        940        950        960 
LKAYVLDDIN LCGYFAYSLS DRSAPKSGFY RYAANQFEPK PSMKHYRRII DSNGFLGSGT 

       970        980        990       1000       1010 
LGRFCPEEYT VCTECGFFQT RKSLLVFISF LVFTFIISLA LIFHYSKKGQ RSYK

O35082 in FASTA format

View entry in original UniProtKB/Swiss-Prot format
View entry in raw text format (no links)
Report form for errors/updates in this UniProtKB/Swiss-Prot entry

	BLAST submission on ExPASy/SIB or at NCBI (USA)		Sequence analysis tools: ProtParam, ProtScale, Compute pI/Mw, PeptideMass, PeptideCutter, Dotlet (Java)
	ScanProsite, MotifScan		Submit a homology modeling request to SWISS-MODEL
	NPSA Sequence analysis tools