Enterokinase (Bovine) Enzyme | TMPRSS15 enzyme

Enterokinase (Bovine) (His tag)

Cat. Num. AAA355467

• Gene Names: TMPRSS15; ENTK; PRSS7
• Purity: > 95% by HPLC
• Synonyms: Enterokinase (Bovine); Enterokinase (Bovine) (His tag); Enteropeptidase; Enterokinase; Serine protease 7; ENTK; TMPRSS15 enzyme

• Host: Pichia pastoris
• Purity/Purification: > 95% by HPLC
• Form/Format: Lyophilized

• Reconstitution: Reconstitute with sterile water, then aliquot the reconstituted enzyme at -20°C.
• Activity: 10,000 U/mg
• Unit Definition: One unit is defined as the amount of enzyme needed to cleave 50 ug of control fusion protein for 16 hours at 20°C in the buffer of 20 mM Tris-HCl, 50 mM NaCl, pH 7.2. (The enzyme cleavage condition will be different for different fusion proteins.)
• Recommended Cleavage Condition: At 20°C in a buffer with 20 mM Tris-HCl, 50 mM NaCl (pH 7.2).
• Provided Assisted Reagents: MBS355467-A: Cleavage Control Protein: 10ug, 1 vial; MBS355467-B: Storage/Reaction Buffer: 2ml (10x), 1 vial; MBS355467-C: Sterile Deionized Water: 1ml, 1 vial; ; Note: The following protocols are for reference ONLY. The enzyme cleavage condition will be adjusted for different fusion proteins, it is recommended to test several trials to determine the optimal Enterokinase concentration and experimental conditions including temperatures, incubation times, etc.
• Example for Enzyme Cleavage: 1. Dilute the Enterokinase and Cleavage Control Protein (provided) with cleavage buffer (20mM Tris-HCl, 50mM NaCl, pH 7.2) to the final concentrations of 0.3mg/ml and 1.0g/ml respectively. ; 2. Mix the related reagents according to the following in a 1.5ml tube: ; - Cleavage Buffer (10x): 5ul; - Target Protein: 20ul; - Diluted Enterkinase: 2ul; - Sterile Deionized Water: 23ul; - Total: 50ul; 3. Incubate at room temperature (20-21°C for 1h, 3h, 6h, and 24h respectively. ; 4. Stop the reaction by storing at -20°C. ; 5. Analyze the cleavage effect of these samples by SDS-PAGE electrophoresis. ; ; Note: The suggested ratio of enzyme and substrate is about 1 U/50ug to 100ug. (The optimal ratio should be determined by end user.)
• Usage of Cleavage Control Protein: 1. Dilute the Enterokinase with 200ul of Sterile Deionized Water, and Cleavage Control Protein with 15ul of Sterile Deionized Water respectively. ; 2. Mix the related reagents according to the following in a 1.5ml tube: ; - Cleavage Control Protein: 15ul; - Diluted Enterkinase: 2ul; - Storage/Reaction Buffer (1x): 17ul; - Total: 34ul; 3. Incubate at room temperature (20-21°C) for 16h.; 4. Add 10ul of 2x concentrated SDS-PAGE buffer into 10ul of samples.; 5. Boil the samples for 5min and analyse the cleavage effect of these samples by SDS-PAGE electrophoresis.
• Preparation and Storage: Upon receipt, store at -80°C for long term.; Avoid repeated freeze and thaw cycles.

Related Product Information for TMPRSS15 enzyme

Description: Recombinant full length bovine Enterokinase with 6 His tags.

Introduction: Recombinant Enterokinase is the light chain of bovine Enterokinase, which is expressed in yeast and purified to yield a high enzyme activity preparation. Enterokinase recognizes the sequence Asp-Asp-Asp-Asp-Lys and cleaves the peptide bond after the lysine residue. This enzyme can be used to cleave any fusion protein that carries this sequence. This product has higher cleavage activity than the native enzymes with 6 His tags which can be deleted after cleavage, and can also removed the rest enzymes rapidly without influencing the cleavage activity.

Background: Enterokinase (also called enteropeptidase, or protease serine 7, PRSS7) is an intestinal enzyme responsible for initiating activation of pancreatic proteolytic proenzymes (trypsin, chymotrypsin and carboxypeptidase A). It is a serine protease consisting of a disulfide-linked 82–140 kDa heavy chain which anchors enterokinase in the intestinal brush border membrane and a 35–62 kDa light chain which contains the catalytic subunit.1 Enteropeptidase is a part of the chymotrypsin-clan of serine proteases, and is structurally similar to these proteins.2 The gene catalyzes the conversion of trypsinogen to trypsin which in turn activates other proenzymes including chymotrypsinogen, procarboxypeptidases, and proelastases.

Product Categories/Family for TMPRSS15 enzyme

Instrumental Enzyme

NCBI and Uniprot Product Information

• NCBI GI #: 223942069
• NCBI GeneID: 5651
• NCBI Accession #: NP_002763.2
• NCBI GenBank Nucleotide #: NM_002772.2
• UniProt Accession #: P98073; Q2NKL7
• NCBI Official Full Name: enteropeptidase
• NCBI Official Synonym Full Names: transmembrane protease, serine 15
• NCBI Official Symbol: TMPRSS15
• NCBI Official Synonym Symbols: ENTK; PRSS7
• NCBI Protein Information: enteropeptidase; enterokinase; serine protease 7; protease, serine, 7 (enterokinase)
• UniProt Protein Name: Enteropeptidase
• Protein Family: Enteropeptidase
• UniProt Gene Name: TMPRSS15
• UniProt Synonym Gene Names: ENTK; PRSS7
• UniProt Entry Name: ENTK_HUMAN

NCBI Description

This gene encodes an enzyme that converts the pancreatic proenzyme trypsinogen to trypsin, which activates other proenzymes including chymotrypsinogen and procarboxypeptidases. The precursor protein is cleaved into two chains that form a heterodimer linked by a disulfide bond. This protein is a member of the trypsin family of peptidases. Mutations in this gene cause enterokinase deficiency, a malabsorption disorder characterized by diarrhea and failure to thrive. [provided by RefSeq, Jul 2008]

Uniprot Description

Function: Responsible for initiating activation of pancreatic proteolytic proenzymes (trypsin, chymotrypsin and carboxypeptidase A). It catalyzes the conversion of trypsinogen to trypsin which in turn activates other proenzymes including chymotrypsinogen, procarboxypeptidases, and proelastases.

Catalytic activity: Activation of trypsinogen by selective cleavage of 6-Lys-|-Ile-7 bond.

Subunit structure: Heterodimer of a catalytic (light) chain and a multidomain (heavy) chain linked by a disulfide bond.

Subcellular location: Membrane; Single-pass type II membrane protein

Probable.

Tissue specificity: Intestinal brush border.

Post-translational modification: The chains are derived from a single precursor that is cleaved by a trypsin-like protease.

Involvement in disease: Enterokinase deficiency (ENTKD) [MIM:226200]: Life-threatening intestinal malabsorption disorder characterized by diarrhea and failure to thrive.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.2

Sequence similarities: Belongs to the peptidase S1 family.Contains 2 CUB domains.Contains 2 LDL-receptor class A domains.Contains 1 MAM domain.Contains 1 peptidase S1 domain.Contains 1 SEA domain.Contains 1 SRCR domain.

Sequence caution: The sequence CAB90389.1 differs from that shown. Reason: Erroneous gene model prediction.