Reviewed, UniProtKB/Swiss-Prot P0C1T0 (MMEL1_RAT)

Names and origin

Protein names

Recommended name:
    Membrane metallo-endopeptidase-like 1
    EC=3.4.24.11
Alternative name(s):
    Neprilysin-2
    Neprilysin II
    NL2
    NEPII
    NEP2(m)
Cleaved into the following chain:
    1- Recommended name:
            Membrane metallo-endopeptidase-like 1, soluble form
        Alternative name(s):
            Neprilysin-2 secreted
              Short name=NEP2(s)

Gene names

Name:	Mmel1
Synonyms:	Mell1, Nep2

Organism

Rattus norvegicus (Rat)

Taxonomic identifier

10116 [NCBI]

Taxonomic lineage

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

Protein attributes

Sequence length	774 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	Metalloprotease involved in sperm function, possibly by modulating the processes of fertilization and early embryonic development. Degrades a broad variety of small peptides with a preference for peptides shorter than 3 kDa containing neutral bulky aliphatic or aromatic amino acid residues. Shares the same substrate specificiy with MME and cleaves peptides at the same amide bond By similarity.
Catalytic activity	Preferential cleavage of polypeptides between hydrophobic residues, particularly with Phe or Tyr at P1'.
Cofactor	Binds 1 zinc ion per subunit By similarity.
Enzyme regulation	Inhibited by thiorphan and phosphoramidon.
Subcellular location	Membrane; Single-pass type II membrane protein. Secreted. Note: A secreted form produced by proteollytic cleavage also exists Probable.
Tissue specificity	Specifically expressed in testis and central nervous system (CNS) (at protein level). Restricted to developing and differentiating fields of the CNS. The soluble form is expressed in round spermatides. Ref.1 Ref.4
Post-translational modification	N-glycosylated. Ref.6
Sequence similarities	Belongs to the peptidase M13 family.
Biophysicochemical properties	Kinetic parameters: KM=62 µM for Tyrosyl-D-Ala(2)-Leu(5)-enkephalin KM=27 µM for Suc-AA-F-AMC (Membrane metallo-endopeptidase-like 1) KM=50 µM for Suc-AA-F-AMC (Membrane metallo-endopeptidase-like 1, soluble form) KM=40 µM for Suc-AA-V-AMC (Membrane metallo-endopeptidase-like 1) KM=100 µM for Suc-AA-V-AMC (Membrane metallo-endopeptidase-like 1, soluble form) KM=23 µM for Suc-G-LF-AMC (Membrane metallo-endopeptidase-like 1) KM=35 µM for Suc-G-LF-AMC (Membrane metallo-endopeptidase-like 1, soluble form)

Ontologies

Keywords
Cellular component	Membrane Secreted
Coding sequence diversity	Alternative splicing
Domain	Coiled coil Signal-anchor Transmembrane
Ligand	Metal-binding Zinc
Molecular function	Hydrolase Metalloprotease Protease
PTM	Disulfide bond Glycoprotein
Gene Ontology (GO)
Biological process	proteolysis Inferred from electronic annotation. Source: InterPro
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell integral to membrane Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	metalloendopeptidase activity Ref.6 Inferred from electronic annotation. Source: InterPro zinc ion binding Ref.5 Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Alternative products

This entry describes 4 isoforms produced by alternative splicing. [Align] [Select]

Isoform 1 (identifier: P0C1T0-1)

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.

Isoform 2 (identifier: P0C1T0-2)

The sequence of this isoform differs from the canonical sequence as follows:
     51-73: Missing.

Isoform 3 (identifier: P0C1T0-3)

The sequence of this isoform differs from the canonical sequence as follows:
     246-267: Missing.

Isoform 4 (identifier: P0C1T0-4)

The sequence of this isoform differs from the canonical sequence as follows:
     303-313: Missing.

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 774	774	Membrane metallo-endopeptidase-like 1		PRO_0000248419
	Chain	73 – 774	702	Membrane metallo-endopeptidase-like 1, soluble form By similarity		PRO_0000248420
Regions
	Topological domain	1 – 26	26	Cytoplasmic Potential
	Transmembrane	27 – 47	21	Signal-anchor for type II membrane protein Potential
	Topological domain	48 – 774	727	Lumenal Potential
	Coiled coil	532 – 558	27	Potential
	Motif	14 – 21	8	Stop-transfer sequence Potential
Sites
	Active site	615	1	By similarity
	Active site	675	1	Proton donor By similarity
	Metal binding	608	1	Zinc; catalytic By similarity
	Metal binding	612	1	Zinc; catalytic By similarity
	Metal binding	671	1	Zinc; catalytic By similarity
	Binding site	131	1	Substrate carboxyl Probable
	Site	72 – 73	2	Cleavage By similarity
Amino acid modifications
	Glycosylation	173	1	N-linked (GlcNAc...) Potential
	Glycosylation	203	1	N-linked (GlcNAc...) Potential
	Glycosylation	239	1	N-linked (GlcNAc...) Potential
	Glycosylation	312	1	N-linked (GlcNAc...) Potential
	Glycosylation	345	1	N-linked (GlcNAc...) Potential
	Glycosylation	633	1	N-linked (GlcNAc...) Potential
	Glycosylation	652	1	N-linked (GlcNAc...) Potential
	Glycosylation	703	1	N-linked (GlcNAc...) Potential
	Disulfide bond	85 ↔ 90		By similarity
	Disulfide bond	108 ↔ 759		By similarity
	Disulfide bond	116 ↔ 719		By similarity
	Disulfide bond	171 ↔ 434		By similarity
	Disulfide bond	645 ↔ 771		By similarity
Natural variations
	Alternative sequence	51 – 73	23	Missing in isoform 2.		VSP_020292
	Alternative sequence	246 – 267	22	Missing in isoform 3.		VSP_020293
	Alternative sequence	303 – 313	11	Missing in isoform 4.		VSP_020294
Experimental info
	Mutagenesis	131	1	R → M: Impairs binding of free carboxylate group of substrates. Ref.5
	Mutagenesis	133	1	S → G: Impairs both substrate-binding and enzyme activity. Ref.5
	Mutagenesis	567	1	N → G: Impairs the enzyme specificity. Ref.5
	Mutagenesis	739	1	L → G: Impairs both substrate-binding and enzyme activity. Ref.5
	Mutagenesis	751	1	P → A: Loss of function. Ref.7
	Mutagenesis	764	1	P → Y: Induces a 3-fold decrease of specific activity. Ref.7
	Mutagenesis	774	1	W → F: No effect. Ref.7

Sequences

	Sequence		Length	Mass (Da)
	Isoform 1 [UniParc]. Last modified September 5, 2006. Version 1. Checksum: F95E2088CA4B64AD	FASTA	774	89,197
	10 20 30 40 50 60 MGKSESSVGM MERADNCGRR RLGFVECGLL VLLTLLLMGA IVTLGVFYSI GKQLPLLNSL 70 80 90 100 110 120 LHVSRHERTV VKRVLRDSSQ KSDICTTPSC VIAAARILQN MDQSKKPCDN FYQYACGGWL 130 140 150 160 170 180 RHHVIPETNS RYSVFDILRD ELEVILKGVL EDSSVQHRPA VEKAKTLYRS CMNQSVIEKR 190 200 210 220 230 240 DSEPLLNVLD MIGGWPVAMD KWNETMGPKW ELERQLAVLN SQFNRRVLID LFIWNDDQNS 250 260 270 280 290 300 SRHVIYIDQP TLGMPSREYY FKEDSHRVRE AYLQFMTSVA TMLRRDLNLP GETDLVQEEM 310 320 330 340 350 360 AQVLHLETHL ANATVPQEKR HDVTALYHRM GLEELQERFG LKGFNWTLFI QNVLSSVQVE 370 380 390 400 410 420 LLPNEEVVVY GIPYLENLEE IIDVFPAQTL QNYLVWRLVL DRIGSLSQRF KEARVDYRKA 430 440 450 460 470 480 LYGTTMEEVR WRECVSYVNS NMESAVGSLY IKRAFSKDSK SIVSELIEKI RSVFVDNLDE 490 500 510 520 530 540 LNWMDEESKK KAQEKALNIR EQIGYPDYIL EDNNRHLDEE YSSLTFSEDL YFENGLQNLK 550 560 570 580 590 600 NNAQRSLKKL REKVDQNLWI IGAAVVNAFY SPNRNLIVFP AGILQPPFFS KDQPQALNFG 610 620 630 640 650 660 GIGMVIGHEI THGFDDNGRN FDKNGNMLDW WSNFSARHFR QQSQCMIYQY SNFSWELADN 670 680 690 700 710 720 QNVNGFSTLG ENIADNGGVR QAYKAYLQWL AEGGRDQRLP GLNLTYAQLF FINYAQVWCG 730 740 750 760 770 SYRPEFAIQS IKTDVHSPLN AQVLGSLQNL PGFSEAFHCP RGSPMHPMNR CRIW
	Isoform 2. Checksum: 1995504DF1EFCDDF	FASTA	751	86,533
	10 20 30 40 50 60 MGKSESSVGM MERADNCGRR RLGFVECGLL VLLTLLLMGA IVTLGVFYSI VLRDSSQKSD 70 80 90 100 110 120 ICTTPSCVIA AARILQNMDQ SKKPCDNFYQ YACGGWLRHH VIPETNSRYS VFDILRDELE 130 140 150 160 170 180 VILKGVLEDS SVQHRPAVEK AKTLYRSCMN QSVIEKRDSE PLLNVLDMIG GWPVAMDKWN 190 200 210 220 230 240 ETMGPKWELE RQLAVLNSQF NRRVLIDLFI WNDDQNSSRH VIYIDQPTLG MPSREYYFKE 250 260 270 280 290 300 DSHRVREAYL QFMTSVATML RRDLNLPGET DLVQEEMAQV LHLETHLANA TVPQEKRHDV 310 320 330 340 350 360 TALYHRMGLE ELQERFGLKG FNWTLFIQNV LSSVQVELLP NEEVVVYGIP YLENLEEIID 370 380 390 400 410 420 VFPAQTLQNY LVWRLVLDRI GSLSQRFKEA RVDYRKALYG TTMEEVRWRE CVSYVNSNME 430 440 450 460 470 480 SAVGSLYIKR AFSKDSKSIV SELIEKIRSV FVDNLDELNW MDEESKKKAQ EKALNIREQI 490 500 510 520 530 540 GYPDYILEDN NRHLDEEYSS LTFSEDLYFE NGLQNLKNNA QRSLKKLREK VDQNLWIIGA 550 560 570 580 590 600 AVVNAFYSPN RNLIVFPAGI LQPPFFSKDQ PQALNFGGIG MVIGHEITHG FDDNGRNFDK 610 620 630 640 650 660 NGNMLDWWSN FSARHFRQQS QCMIYQYSNF SWELADNQNV NGFSTLGENI ADNGGVRQAY 670 680 690 700 710 720 KAYLQWLAEG GRDQRLPGLN LTYAQLFFIN YAQVWCGSYR PEFAIQSIKT DVHSPLNAQV 730 740 750 LGSLQNLPGF SEAFHCPRGS PMHPMNRCRI W
	Isoform 3. Checksum: A7E932880187EFFE	FASTA	752	86,482
	10 20 30 40 50 60 MGKSESSVGM MERADNCGRR RLGFVECGLL VLLTLLLMGA IVTLGVFYSI GKQLPLLNSL 70 80 90 100 110 120 LHVSRHERTV VKRVLRDSSQ KSDICTTPSC VIAAARILQN MDQSKKPCDN FYQYACGGWL 130 140 150 160 170 180 RHHVIPETNS RYSVFDILRD ELEVILKGVL EDSSVQHRPA VEKAKTLYRS CMNQSVIEKR 190 200 210 220 230 240 DSEPLLNVLD MIGGWPVAMD KWNETMGPKW ELERQLAVLN SQFNRRVLID LFIWNDDQNS 250 260 270 280 290 300 SRHVIVREAY LQFMTSVATM LRRDLNLPGE TDLVQEEMAQ VLHLETHLAN ATVPQEKRHD 310 320 330 340 350 360 VTALYHRMGL EELQERFGLK GFNWTLFIQN VLSSVQVELL PNEEVVVYGI PYLENLEEII 370 380 390 400 410 420 DVFPAQTLQN YLVWRLVLDR IGSLSQRFKE ARVDYRKALY GTTMEEVRWR ECVSYVNSNM 430 440 450 460 470 480 ESAVGSLYIK RAFSKDSKSI VSELIEKIRS VFVDNLDELN WMDEESKKKA QEKALNIREQ 490 500 510 520 530 540 IGYPDYILED NNRHLDEEYS SLTFSEDLYF ENGLQNLKNN AQRSLKKLRE KVDQNLWIIG 550 560 570 580 590 600 AAVVNAFYSP NRNLIVFPAG ILQPPFFSKD QPQALNFGGI GMVIGHEITH GFDDNGRNFD 610 620 630 640 650 660 KNGNMLDWWS NFSARHFRQQ SQCMIYQYSN FSWELADNQN VNGFSTLGEN IADNGGVRQA 670 680 690 700 710 720 YKAYLQWLAE GGRDQRLPGL NLTYAQLFFI NYAQVWCGSY RPEFAIQSIK TDVHSPLNAQ 730 740 750 VLGSLQNLPG FSEAFHCPRG SPMHPMNRCR IW
	Isoform 4. Checksum: 60A7053E8AE2D62A	FASTA	763	87,997
	10 20 30 40 50 60 MGKSESSVGM MERADNCGRR RLGFVECGLL VLLTLLLMGA IVTLGVFYSI GKQLPLLNSL 70 80 90 100 110 120 LHVSRHERTV VKRVLRDSSQ KSDICTTPSC VIAAARILQN MDQSKKPCDN FYQYACGGWL 130 140 150 160 170 180 RHHVIPETNS RYSVFDILRD ELEVILKGVL EDSSVQHRPA VEKAKTLYRS CMNQSVIEKR 190 200 210 220 230 240 DSEPLLNVLD MIGGWPVAMD KWNETMGPKW ELERQLAVLN SQFNRRVLID LFIWNDDQNS 250 260 270 280 290 300 SRHVIYIDQP TLGMPSREYY FKEDSHRVRE AYLQFMTSVA TMLRRDLNLP GETDLVQEEM 310 320 330 340 350 360 AQTVPQEKRH DVTALYHRMG LEELQERFGL KGFNWTLFIQ NVLSSVQVEL LPNEEVVVYG 370 380 390 400 410 420 IPYLENLEEI IDVFPAQTLQ NYLVWRLVLD RIGSLSQRFK EARVDYRKAL YGTTMEEVRW 430 440 450 460 470 480 RECVSYVNSN MESAVGSLYI KRAFSKDSKS IVSELIEKIR SVFVDNLDEL NWMDEESKKK 490 500 510 520 530 540 AQEKALNIRE QIGYPDYILE DNNRHLDEEY SSLTFSEDLY FENGLQNLKN NAQRSLKKLR 550 560 570 580 590 600 EKVDQNLWII GAAVVNAFYS PNRNLIVFPA GILQPPFFSK DQPQALNFGG IGMVIGHEIT 610 620 630 640 650 660 HGFDDNGRNF DKNGNMLDWW SNFSARHFRQ QSQCMIYQYS NFSWELADNQ NVNGFSTLGE 670 680 690 700 710 720 NIADNGGVRQ AYKAYLQWLA EGGRDQRLPG LNLTYAQLFF INYAQVWCGS YRPEFAIQSI 730 740 750 760 KTDVHSPLNA QVLGSLQNLP GFSEAFHCPR GSPMHPMNRC RIW

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Neprilysin II: a putative novel metalloprotease and its isoforms in CNS and testis." Ouimet T., Facchinetti P., Rose C., Bonhomme M.-C., Gros C., Schwartz J.-C. Biochem. Biophys. Res. Commun. 271:565-570(2000) [PubMed: 10814502] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS 1; 2 AND 3), TISSUE SPECIFICITY.
[2]	Erratum Ouimet T., Facchinetti P., Rose C., Bonhomme M.-C., Gros C., Schwartz J.-C. Biochem. Biophys. Res. Commun. 275:247-247(2000)
[3]	"Genome sequence of the Brown Norway rat yields insights into mammalian evolution." Gibbs R.A., Weinstock G.M., Metzker M.L., Muzny D.M., Sodergren E.J., Scherer S., Scott G., Steffen D., Worley K.C., Burch P.E., Okwuonu G., Hines S., Lewis L., Deramo C., Delgado O., Dugan-Rocha S., Miner G., Morgan M. , Hawes A., Gill R., Holt R.A., Adams M.D., Amanatides P.G., Baden-Tillson H., Barnstead M., Chin S., Evans C.A., Ferriera S., Fosler C., Glodek A., Gu Z., Jennings D., Kraft C.L., Nguyen T., Pfannkoch C.M., Sitter C., Sutton G.G., Venter J.C., Woodage T., Smith D., Lee H.-M., Gustafson E., Cahill P., Kana A., Doucette-Stamm L., Weinstock K., Fechtel K., Weiss R.B., Dunn D.M., Green E.D., Blakesley R.W., Bouffard G.G., De Jong P.J., Osoegawa K., Zhu B., Marra M., Schein J., Bosdet I., Fjell C., Jones S., Krzywinski M., Mathewson C., Siddiqui A., Wye N., McPherson J., Zhao S., Fraser C.M., Shetty J., Shatsman S., Geer K., Chen Y., Abramzon S., Nierman W.C., Havlak P.H., Chen R., Durbin K.J., Egan A., Ren Y., Song X.-Z., Li B., Liu Y., Qin X., Cawley S., Cooney A.J., D'Souza L.M., Martin K., Wu J.Q., Gonzalez-Garay M.L., Jackson A.R., Kalafus K.J., McLeod M.P., Milosavljevic A., Virk D., Volkov A., Wheeler D.A., Zhang Z., Bailey J.A., Eichler E.E., Tuzun E., Birney E., Mongin E., Ureta-Vidal A., Woodwark C., Zdobnov E., Bork P., Suyama M., Torrents D., Alexandersson M., Trask B.J., Young J.M., Huang H., Wang H., Xing H., Daniels S., Gietzen D., Schmidt J., Stevens K., Vitt U., Wingrove J., Camara F., Mar Alba M., Abril J.F., Guigo R., Smit A., Dubchak I., Rubin E.M., Couronne O., Poliakov A., Huebner N., Ganten D., Goesele C., Hummel O., Kreitler T., Lee Y.-A., Monti J., Schulz H., Zimdahl H., Himmelbauer H., Lehrach H., Jacob H.J., Bromberg S., Gullings-Handley J., Jensen-Seaman M.I., Kwitek A.E., Lazar J., Pasko D., Tonellato P.J., Twigger S., Ponting C.P., Duarte J.M., Rice S., Goodstadt L., Beatson S.A., Emes R.D., Winter E.E., Webber C., Brandt P., Nyakatura G., Adetobi M., Chiaromonte F., Elnitski L., Eswara P., Hardison R.C., Hou M., Kolbe D., Makova K., Miller W., Nekrutenko A., Riemer C., Schwartz S., Taylor J., Yang S., Zhang Y., Lindpaintner K., Andrews T.D., Caccamo M., Clamp M., Clarke L., Curwen V., Durbin R.M., Eyras E., Searle S.M., Cooper G.M., Batzoglou S., Brudno M., Sidow A., Stone E.A., Payseur B.A., Bourque G., Lopez-Otin C., Puente X.S., Chakrabarti K., Chatterji S., Dewey C., Pachter L., Bray N., Yap V.B., Caspi A., Tesler G., Pevzner P.A., Haussler D., Roskin K.M., Baertsch R., Clawson H., Furey T.S., Hinrichs A.S., Karolchik D., Kent W.J., Rosenbloom K.R., Trumbower H., Weirauch M., Cooper D.N., Stenson P.D., Ma B., Brent M., Arumugam M., Shteynberg D., Copley R.R., Taylor M.S., Riethman H., Mudunuri U., Peterson J., Guyer M., Felsenfeld A., Old S., Mockrin S., Collins F.S. Nature 428:493-521(2004) [PubMed: 15057822] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Brown Norway.
[4]	"Ontogeny, regional and cellular distribution of the novel metalloprotease neprilysin 2 in the rat: a comparison with neprilysin and endothelin-converting enzyme-1." Facchinetti P., Rose C., Schwartz J.C., Ouimet T. Neuroscience 118:627-639(2003) [PubMed: 12710972] [Abstract] Cited for: TISSUE SPECIFICITY.
[5]	"A three-dimensional model of the neprilysin 2 active site based on the X-ray structure of neprilysin. Identification of residues involved in substrate hydrolysis and inhibitor binding of neprilysin 2." Voisin S., Rognan D., Gros C., Ouimet T. J. Biol. Chem. 279:46172-46181(2004) [PubMed: 15294904] [Abstract] Cited for: BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ARG-131; SER-133; ASN-567 AND LEU-739.
[6]	"Cell-specific activity of neprilysin 2 isoforms and enzymic specificity compared with neprilysin." Rose C., Voisin S., Gros C., Schwartz J.-C., Ouimet T. Biochem. J. 363:697-705(2002) [PubMed: 11964170] [Abstract] Cited for: GLYCOSYLATION, BIOPHYSICOCHEMICAL PROPERTIES.
[7]	"The ultimate tryptophan residue of neprilysin 2 is not involved in protein maturation and enzymatic activity." Voisin S., Ouimet T. Biochem. Biophys. Res. Commun. 335:356-360(2005) [PubMed: 16081046] [Abstract] Cited for: MUTAGENESIS OF PRO-751; PRO-764 AND TRP-774.
+	Additional computationally mapped references.

Cross-references

Sequence databases
EMBL GenBank DDBJ	AABR03040205 Genomic DNA. No translation available.
IPI	IPI00785587. IPI00785597. IPI00785603. IPI00869674.
UniGene	Rn.50873
3D structure databases
SMR	P0C1T0. Positions 83-774.
ModBase	Search...
Genome annotation databases
Ensembl	ENSRNOT00000017957; ENSRNOP00000017957; ENSRNOG00000012593; Rattus norvegicus. [Genome view]
NMPDR	fig|10116.3.peg.24529.
Organism-specific databases
RGD	1309299. Mell1.
Phylogenomic databases
eggNOG	roNOG11771.
HOVERGEN	P0C1T0.
Enzyme and pathway databases
BRENDA	3.4.24.11. 248.
Gene expression databases
ArrayExpress	P0C1T0.
Genevestigator	P0C1T0.
Family and domain databases
InterPro	IPR000718. Peptidase_M13. IPR018497. Peptidase_M13_C. IPR008753. Peptidase_M13_N. [Graphical view]
PANTHER	PTHR11733. Peptidase_M13. 1 hit.
Pfam	PF01431. Peptidase_M13. 1 hit. PF05649. Peptidase_M13_N. 1 hit. [Graphical view]
PRINTS	PR00786. NEPRILYSIN.
PROSITE	PS00142. ZINC_PROTEASE. 1 hit. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

MMEL1_RAT

Accession

Primary (citable) accession number: P0C1T0

Entry history

Integrated into UniProtKB/Swiss-Prot:	September 5, 2006
Last sequence update:	September 5, 2006
Last modified:	January 19, 2010
This is version 31 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HPI (Human Proteome Initiative)