ENPP-2
Human Recombinant ENPP2 - His Tagged
Reference ID:KB-3922
Western Blot
ELISA
FACS
Protein:Protein
Flow Cytometry
Gene of Interest
Gene Synonyms:ENPP2;ATX;PDNP2
Protein Names:Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 (E-NPP 2) (EC 3.1.4.39) (Autotaxin) (Extracellular lysophospholipase D) (LysoPLD)
Accession Data
Organism:Homo sapiens (Human)
Mass (kDa):989.94
Length (aa):863
Sequence:MARRSSFQSCQIISLFTFAVGVNICLGFTAHRIKRAEGWEEGPPTVLSDSPWTNISGSCKGRCFELQEAGPPDCRCDNLCKSYTSCCHDFDELCLKTARGWECTKDRCGEVRNEENACHCSEDCLARGDCCTNYQVVCKGESHWVDDDCEEIKAAECPAGFVRPPLIIFSVDGFRASYMKKGSKVMPNIEKLRSCGTHSPYMRPVYPTKTFPNLYTLATGLYPESHGIVGNSMYDPVFDATFHLRGREKFNHRWWGGQPLWITATKQGVKAGTFFWSVVIPHERRILTILQWLTLPDHERPSVYAFYSEQPDFSGHKYGPFGPEMTNPLREIDKIVGQLMDGLKQLKLHRCVNVIFVGDHGMEDVTCDRTEFLSNYLTNVDDITLVPGTLGRIRSKFSNNAKYDPKAIIANLTCKKPDQHFKPYLKQHLPKRLHYANNRRIEDIHLLVERRWHVARKPLDVYKKPSGKCFFQGDHGFDNKVNSMQTVFVGYGSTFKYKTKVPPFENIELYNVMCDLLGLKPAPNNGTHGSLNHLLRTNTFRPTMPEEVTRPNYPGIMYLQSDFDLGCTCDDKVEPKNKLDELNKRLHTKGSTEERHLLYGRPAVLYRTRYDILYHTDFESGYSEIFLMPLWTSYTVSKQAEVSSVPDHLTSCVRPDVRVSPSFSQNCLAYKNDKQMSYGFLFPPYLSSSPEAKYDAFLVTNMVPMYPAFKRVWNYFQRVLVKKYASERNGVNVISGPIFDYDYDGLHDTEDKIKQYVEGSSIPVPTHYYSIITSCLDFTQPADKCDGPLSVSSFILPHRPDNEESCNSSEDESKWVEELMKMHTARVRDIEHLTSLDFFRKTSRSYPEILTLKTYLHTYESEI
Proteomics (Proteome ID):UP000005640
Proteomics (Chromosome): Chromosome 8
Active Site:ACT_SITE 210 210 Nucleophile. {ECO:0000250|UniProtKB:Q9R1E6}.
Activity Regulation: Inhibited by lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P). Inhibited by EDTA and EGTA (Probable). {ECO:0000305|PubMed:15769751, ECO:0000305|PubMed:18175805, ECO:0000305|PubMed:21240271}.
Binding Site:BINDING 231 231 Substrate. {ECO:0000244|PDB:4ZG7, ECO:0000305|PubMed:26371182}.; BINDING 307 307 Substrate. {ECO:0000250|UniProtKB:Q9R1E6}.
Catalytic Activity: Reaction=1-O-alkyl-sn-glycero-3-phosphoethanolamine + H2O = 1-O-alkyl-sn-glycero-3-phosphate + ethanolamine + H(+); Xref=Rhea:RHEA:15965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:57603, ChEBI:CHEBI:58014, ChEBI:CHEBI:76168; EC=3.1.4.39; Evidence={ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:15769751, ECO:0000269|PubMed:18175805, ECO:0000269|PubMed:21240271, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931};
Cofactor:Name=Zn(2+); Xref=ChEBI:CHEBI:29105; Evidence={ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931, ECO:0000269|PubMed:28414242}; Note=Binds 2 Zn(2+) ions per subunit. {ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931, ECO:0000269|PubMed:28414242}; Name=Ca(2+); Xref=ChEBI:CHEBI:29108; Evidence={ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931, ECO:0000269|PubMed:28414242}; Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931, ECO:0000269|PubMed:28414242};
Function [CC]:Hydrolyzes lysophospholipids to produce the signaling molecule lysophosphatidic acid (LPA) in extracellular fluids (PubMed:15769751, PubMed:26371182, PubMed:27754931). Major substrate is lysophosphatidylcholine (PubMed:12176993, PubMed:27754931). Also can act on sphingosylphosphorylcholine producing sphingosine-1-phosphate, a modulator of cell motility. Can hydrolyze, in vitro, bis-pNPP, to some extent pNP-TMP, and barely ATP (PubMed:15769751, PubMed:12176993). Involved in several motility-related processes such as angiogenesis and neurite outgrowth. Acts as an angiogenic factor by stimulating migration of smooth muscle cells and microtubule formation (PubMed:11559573). Stimulates migration of melanoma cells, probably via a pertussis toxin-sensitive G protein (PubMed:1733949). May have a role in induction of parturition (PubMed:12176993). Possible involvement in cell proliferation and adipose tissue development (Probable). Tumor cell motility-stimulating factor (PubMed:1733949, PubMed:11559573). {ECO:0000269|PubMed:11559573, ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:15769751, ECO:0000269|PubMed:1733949, ECO:0000269|PubMed:21240271, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931, ECO:0000305|PubMed:15700135}.
Kinetics:BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.5 mM for 16:0-LPC (at pH 8.5) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; KM=5.5 mM for pNP-TMP (at pH 8.5) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; KM=11.3 mM for pNppp (isoform 1) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; KM=5.7 mM for pNppp (isoform 2) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; KM=19.8 mM for pNppp (isoform 3) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; Vmax=1.9 nmol/min/ug enzyme with pNppp as substrate (isoform 1) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; Vmax=0.67 nmol/min/ug enzyme with pNppp as substrate (isoform 2) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805}; Vmax=1.6 nmol/min/ug enzyme with pNppp as substrate (isoform 3) {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805};
Metal Binding:METAL 172 172 Zinc 1; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 210 210 Zinc 1; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 312 312 Zinc 2; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000269|PubMed:26371182}.; METAL 316 316 Zinc 2; via tele nitrogen; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000269|PubMed:26371182}.; METAL 359 359 Zinc 1; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 360 360 Zinc 1; via tele nitrogen; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 475 475 Zinc 2; via tele nitrogen; catalytic. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000269|PubMed:26371182}.; METAL 740 740 Calcium. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 742 742 Calcium. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 744 744 Calcium. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000269|PubMed:26371182}.; METAL 746 746 Calcium; via carbonyl oxygen. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.; METAL 748 748 Calcium. {ECO:0000244|PDB:4ZG6, ECO:0000244|PDB:4ZG7, ECO:0000244|PDB:4ZG9, ECO:0000244|PDB:4ZGA, ECO:0000244|PDB:5KXA, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:27754931}.
Site:SITE 853 853 Essential for catalytic activity. {ECO:0000250|UniProtKB:Q9R1E6}.
Temperature Dependence:BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Isoform 1 and isoform 3 are active from 45 to 60 degrees Celsius. {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805};
Induction:Up-regulated in massively obese subjects with glucose intolerance, and during adipogenesis. {ECO:0000269|PubMed:15700135}.
Tissue Specificity:Detected in blood plasma (at protein level) (PubMed:12176993, PubMed:26371182). Predominantly expressed in brain, placenta, ovary, and small intestine. Expressed in a number of carcinomas such as hepatocellular and prostate carcinoma, neuroblastoma and non-small-cell lung cancer. Expressed in body fluids such as plasma, cerebral spinal fluid (CSF), saliva, follicular and amniotic fluids. Not detected in leukocytes. Isoform 1 is more highly expressed in peripheral tissues than in the central nervous system (CNS). Adipocytes only express isoform 1. Isoform 3 is more highly expressed in the brain than in peripheral tissues. {ECO:0000269|PubMed:12176993, ECO:0000269|PubMed:18175805, ECO:0000269|PubMed:26371182, ECO:0000269|PubMed:8579579, ECO:0000269|PubMed:8586446}.
Chemical Profile | ChEBI:H2O [CHEBI:15377]; Zn(2+) [CHEBI:29105]; H(+) [CHEBI:15378]; ethanolamine [CHEBI:57603]; Ca(2+) [CHEBI:29108]; 1-O-alkyl-sn-glycero-3-phosphoethanolamine [CHEBI:76168]; 1-O-alkyl-sn-glycero-3-phosphate [CHEBI:58014]
ChEBI (Catalytic Activity):H2O [CHEBI:15377]; H(+) [CHEBI:15378]; ethanolamine [CHEBI:57603]; 1-O-alkyl-sn-glycero-3-phosphoethanolamine [CHEBI:76168]; 1-O-alkyl-sn-glycero-3-phosphate [CHEBI:58014]
ChEBI (Cofactor):Zn(2+) [CHEBI:29105]; Ca(2+) [CHEBI:29108]
Mutagenesis:MUTAGEN 170 170 S->E: Reduces lysophospholipase activity by about 70%.; MUTAGEN 210 210 T->A: Loss of lysophospholipase activity. {ECO:0000269|PubMed:21240271}.; MUTAGEN 211 211 F->Y: Reduces lysophospholipase activity by about 70%. {ECO:0000269|PubMed:21240271}.; MUTAGEN 218 218 A->V: Reduces lysophospholipase activity by about 50%. {ECO:0000269|PubMed:21240271}.; MUTAGEN 231 231 N->A: Strongly reduced lysophospholipase activity. {ECO:0000269|PubMed:21240271}.; MUTAGEN 307 307 Y->Q: Reduces lysophospholipase activity by about 70%. {ECO:0000269|PubMed:21240271}.
Reagent Data
Name:ENPP-2
Subcategory:Enzyme
Region:Asp 49 - Ile 863
Molecular Weight:94.5
Source:HEK293
Species:Human
Tag:His
Format:Lyophilized
Purification System:Chromatography
Formulation:Sterile-filtered colorless solution
Formulation Concentration:1 mg/ml
Buffer Volume:Standard
Buffer Solution:PBS
pH:7.4-7.5
Toxicity
Endotoxin Level:< 1%
Aggregate Tested By:SDS-PAGE
Endotoxin Screened:< 0.1 ng/ug
Purity:> 98%
Determined: SDS-PAGE
Stain:Blue
Chromotography:Anion-exchange
Validated: RP-HPLC
Sample Handling
Storage:-20°C
Stability:This bioreagent is stable at 4°C (short-term) and -70°C(long-term). After reconstitution, sample may be stored at 4°C for 2-7 days and below -18°C for future use.
Preparation:Reconstitute in sterile distilled H2O to no less than 100 ug/ml; dilute reconstituted stock further in other aqueous solutions if needed. Please review COA for lot-specific instructions. Final measurements should be determined by the end-user for optimal performance.