Reviewed, UniProtKB/Swiss-Prot O35082 (KLOT_MOUSE)

Names and origin

Protein names

Recommended name:
    Klotho
    EC=3.2.1.31
Cleaved into the following chain:
    1- Recommended name:
            Klotho peptide

Gene names

Name:

Kl

Organism

Mus musculus (Mouse)

Taxonomic identifier

10090 [NCBI]

Taxonomic lineage

Eukaryota › Metazoa › Chordata › Craniata › Vertebrata › Euteleostomi › Mammalia › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Muridae › Murinae › Mus

Protein attributes

Sequence length	1014 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level.

General annotation (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. Ref.1 Ref.4 Ref.5 Ref.8 Ref.13 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. Ref.1 Ref.4 Ref.5 Ref.8 Ref.13
Catalytic activity	A beta-D-glucuronoside + H(2)O = D-glucuronate + an alcohol. Ref.13 Ref.12
Enzyme regulation	Inhibited by D-saccharic acid 1,4-lactone and taurocholic acid. Ref.12
Subunit structure	Homodimer. Ref.11
Subcellular location	Isoform 1: Cell membrane; Single-pass type I membrane protein. Note= Isoform 1 shedding leads to a soluble peptide. Ref.1 Ref.13 Ref.11 Ref.3 Ref.7 Ref.10 Isoform 2: Secreted. Ref.1 Ref.13 Ref.11 Ref.3 Ref.7 Ref.10 Klotho peptide: Secreted. Ref.1 Ref.13 Ref.11 Ref.3 Ref.7 Ref.10
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. Ref.1 Ref.13 Ref.11 Ref.3 Ref.7 Ref.10 Ref.2 Ref.6
Developmental stage	Not expressed in the embryo. Expressed in the kidney of newborns. Ref.1
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. Ref.8 Ref.6 Ref.9
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.
Post-translational modification	N-glycosylated. Ref.11
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.
Sequence similarities	Belongs to the glycosyl hydrolase 1 family. Klotho subfamily.
Biophysicochemical properties	Kinetic parameters: KM=0.249 mM for 4-methylumbelliferylglucuronide KM=0.251 mM for estrone 3-beta-D-glucuronide KM=0.174 mM for beta-estradiol 3-beta-D-glucuronide KM=0.251 mM for estriol 3-beta-D-glucuronide Vmax=0.62 µM/h/µg enzyme pH dependence: Optimum pH is 5.5.

Ontologies

Keywords
Cellular component	Cell membrane Membrane Secreted
Coding sequence diversity	Alternative splicing
Domain	Repeat Signal Transmembrane
Molecular function	Glycosidase Hydrolase
PTM	Glycoprotein Phosphoprotein
Gene Ontology (GO)
Biological process	aging Ref.1 Inferred from mutant phenotype. Source: MGI carbohydrate metabolic process Inferred from electronic annotation. Source: InterPro energy reserve metabolic process Ref.4 Inferred from mutant phenotype. Source: MGI
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-KW integral to membrane Inferred from electronic annotation. Source: UniProtKB-KW plasma membrane Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	beta-glucuronidase activity Inferred from electronic annotation. Source: EC cation binding Inferred from electronic annotation. Source: InterPro protein binding Inferred from physical interaction. Source: IntAct
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
Wnt1	P04426	1	EBI-1570828,EBI-1570911
Wnt3	P17553	1	EBI-1570828,EBI-1570853
Wnt4	P22724	1	EBI-1570828,EBI-1570945
Wnt5a	P22725	1	EBI-1570828,EBI-1570983

Alternative products

This entry describes 2 isoforms produced by alternative splicing. [Align] [Select]

Isoform 1 (identifier: O35082-1)

Also known as: Membrane-bound;

This isoform has been chosen as the 'canonical' sequence. All positional information in this entry refers to it. This is also the sequence that appears in the downloadable versions of the entry.

Notes: Isoform 1 shedding leads to a soluble peptide. Ref.1 Ref.13 Ref.11 Ref.3 Ref.7 Ref.10 Predominates over the secreted form by more than 10 times in all tissues examined.

Isoform 2 (identifier: O35082-2)

Also known as: Secreted;

The sequence of this isoform differs from the canonical sequence as follows:
     537-550: DTTLSQFTDPNVYL → SPLTKPSVGLLLPH
     551-1014: Missing.

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Signal peptide	1 – 34	34	Potential
	Chain	35 – 1014	980	Klotho		PRO_0000042247
	Chain	35 – ?		Klotho peptide		PRO_0000042248
Regions
	Topological domain	35 – 982	948	Extracellular Potential
	Transmembrane	983 – 1003	21	Potential
	Topological domain	1004 – 1014	11	Cytoplasmic Potential
	Region	59 – 508	450	Glycosyl hydrolase-1 1
	Region	517 – 955	439	Glycosyl hydrolase-1 2
Amino acid modifications
	Modified residue	499	1	Phosphotyrosine Ref.14
	Glycosylation	161	1	N-linked (GlcNAc...) Potential
	Glycosylation	285	1	N-linked (GlcNAc...) Potential
	Glycosylation	346	1	N-linked (GlcNAc...) Potential
	Glycosylation	609	1	N-linked (GlcNAc...) Potential
	Glycosylation	614	1	N-linked (GlcNAc...) Potential
	Glycosylation	696	1	N-linked (GlcNAc...) Potential
Natural variations
	Alternative sequence	537 – 550	14	DTTLS…PNVYL → SPLTKPSVGLLLPH in isoform 2.		VSP_015828
	Alternative sequence	551 – 1014	464	Missing in isoform 2.		VSP_015829
Experimental info
	Sequence conflict	854	1	L → V in BAA25308. Ref.2

Sequences

	Sequence		Length	Mass (Da)
	Isoform 1 (Membrane-bound) [UniParc]. Last modified January 1, 1998. Version 1. Checksum: 24BD600D2F969FDC	FASTA	1,014	116,426
	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
	Isoform 2 (Secreted). Checksum: E59AF8F24871BDC8		550	62,348
	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 DNYVQVSPLT 550 KPSVGLLLPH

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Mutation of the mouse klotho gene leads to a syndrome resembling ageing." 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. Nature 390:45-51(1997) [PubMed: 9363890] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS 1 AND 2), FUNCTION, TISSUE SPECIFICITY, DEVELOPMENTAL STAGE, SUBCELLULAR LOCATION. Tissue: Kidney.
[2]	"Structure of the mouse klotho gene and its two transcripts encoding membrane and secreted protein." Shiraki-Iida T., Aizawa H., Matsumura Y., Sekine S., Iida A., Anazawa H., Nagai R., Kuro-o M., Nabeshima Y. FEBS Lett. 424:6-10(1998) [PubMed: 9537505] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], ALTERNATIVE SPLICING (ISOFORMS 1 AND 2), TISSUE SPECIFICITY.
[3]	"Establishment of the anti-Klotho monoclonal antibodies and detection of Klotho protein in kidneys." 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. Biochem. Biophys. Res. Commun. 267:597-602(2000) [PubMed: 10631108] [Abstract] Cited for: TISSUE SPECIFICITY, SUBCELLULAR LOCATION.
[4]	"Disruption of klotho gene causes an abnormal energy homeostasis in mice." Mori K., Yahata K., Mukoyama M., Suganami T., Makino H., Nagae T., Masuzaki H., Ogawa Y., Sugawara A., Nabeshima Y., Nakao K. Biochem. Biophys. Res. Commun. 278:665-670(2000) [PubMed: 11095966] [Abstract] Cited for: FUNCTION.
[5]	"Decreased insulin production and increased insulin sensitivity in the klotho mutant mouse, a novel animal model for human aging." 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. Metabolism 49:1118-1123(2000) [PubMed: 11016890] [Abstract] Cited for: FUNCTION.
[6]	"Upregulation of the klotho gene expression by thyroid hormone and during adipose differentiation in 3T3-L1 adipocytes." Mizuno I., Takahashi Y., Okimura Y., Kaji H., Chihara K. Life Sci. 68:2917-2923(2001) [PubMed: 11411791] [Abstract] Cited for: INDUCTION, TISSUE SPECIFICITY.
[7]	"Expression of Klotho protein in the inner ear." Kamemori M., Ohyama Y., Kurabayashi M., Takahashi K., Nagai R., Furuya N. Hear. Res. 171:103-110(2002) [PubMed: 12204354] [Abstract] Cited for: SUBCELLULAR LOCATION, TISSUE SPECIFICITY.
[8]	"Klotho, a gene related to a syndrome resembling human premature aging, functions in a negative regulatory circuit of vitamin D endocrine system." Tsujikawa H., Kurotaki Y., Fujimori T., Fukuda K., Nabeshima Y. Mol. Endocrinol. 17:2393-2403(2003) [PubMed: 14528024] [Abstract] Cited for: FUNCTION, INDUCTION.
[9]	"HMG-CoA reductase inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells." Narumiya H., Sasaki S., Kuwahara N., Irie H., Kusaba T., Kameyama H., Tamagaki K., Hatta T., Takeda K., Matsubara H. Cardiovasc. Res. 64:331-336(2004) [PubMed: 15485693] [Abstract] Cited for: INDUCTION.
[10]	"Immunohistochemical localization of Klotho protein in brain, kidney, and reproductive organs of mice." Li S.-A., Watanabe M., Yamada H., Nagai A., Kinuta M., Takei K. Cell Struct. Funct. 29:91-99(2004) [PubMed: 15665504] [Abstract] Cited for: SUBCELLULAR LOCATION, TISSUE SPECIFICITY.
[11]	"Secreted Klotho protein in sera and CSF: implication for post-translational cleavage in release of Klotho protein from cell membrane." Imura A., Iwano A., Tohyama O., Tsuji Y., Nozaki K., Hashimoto N., Fujimori T., Nabeshima Y. FEBS Lett. 565:143-147(2004) [PubMed: 15135068] [Abstract] Cited for: CLEAVAGE, TISSUE SPECIFICITY, SUBUNIT, GLYCOSYLATION, SUBCELLULAR LOCATION.
[12]	"Klotho is a novel beta-glucuronidase capable of hydrolyzing steroid beta-glucuronides." Tohyama O., Imura A., Iwano A., Freund J.-N., Henrissat B., Fujimori T., Nabeshima Y. J. Biol. Chem. 279:9777-9784(2004) [PubMed: 14701853] [Abstract] Cited for: ENZYME ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES.
[13]	"Suppression of aging in mice by the hormone Klotho." 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. Science 309:1829-1833(2005) [PubMed: 16123266] [Abstract] Cited for: FUNCTION, LACK OF ENZYMATIC ACTIVITY, TISSUE SPECIFICITY, SUBCELLULAR LOCATION.
[14]	"Qualitative and quantitative analyses of protein phosphorylation in naive and stimulated mouse synaptosomal preparations." 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. Mol. Cell. Proteomics 6:283-293(2007) [PubMed: 17114649] [Abstract] Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT TYR-499, MASS SPECTROMETRY. Tissue: Brain cortex.
+	Additional computationally mapped references.

Web resources

Protein Spotlight

The thread of life - Issue 65 of December 2005

Cross-references

Sequence databases
EMBLGenBankDDBJ	AB005141 mRNA. Translation: BAA23381.1. AB010088 mRNA. Translation: BAA25307.1. AB010091 Genomic DNA. Translation: BAA25308.1. AB010091 Genomic DNA. Translation: BAA25309.1.
RefSeq	NP_038851.1.
UniGene	Mm.6500
3D structure databases
HSSP	HSSP built from PDB template 1OIF based on UniProtKB Q08638.
ModBase	Search...
Protein-protein interaction databases
IntAct	O35082. 4 interactions.
PTM databases
PhosphoSite	O35082.
Genome annotation databases
Ensembl	ENSMUSG00000058488. Mus musculus. [Contig view]
GeneID	16591.
KEGG	mmu:16591.
Organism-specific databases
MGI	MGI:1101771. Kl.
Phylogenomic databases
HOGENOM	O35082.
HOVERGEN	O35082.
Gene expression databases
ArrayExpress	O35082.
CleanEx	MM_KL.
GermOnline	ENSMUSG00000058488. Mus musculus.
Family and domain databases
InterPro	IPR001360. Glyco_hydro_1. IPR013781. Glyco_hydro_sub_cat. [Graphical view]
Gene3D	G3DSA:3.20.20.80. Glyco_hydro_cat. 2 hits.
PANTHER	PTHR10353. Glyco_hydro_1. 1 hit.
Pfam	PF00232. Glyco_hydro_1. 3 hits. [Graphical view]
PRINTS	PR00131. GLHYDRLASE1.
ProDom	PD000650. Glyco_hydro_euk. 2 hits. [Graphical view] [Entries sharing at least one domain]
PROSITE	PS00572. GLYCOSYL_HYDROL_F1_1. False negative. PS00653. GLYCOSYL_HYDROL_F1_2. 1 hit. [Graphical view]
ProtoNet	Search...
Other Resources
NextBio	290141.
SOURCE	Search...

Entry information

Entry name

KLOT_MOUSE

Accession

Primary (citable) accession number: O35082
Secondary accession number(s): O70175, O70621

Entry history

Integrated into UniProtKB/Swiss-Prot:	October 11, 2005
Last sequence update:	January 1, 1998
Last modified:	December 16, 2008
This is version 66 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HPI (Human Proteome Initiative)