ProMMP-13 Recombinant Procollagenase-3

Key Applications	Entrez Gene Number	Species	Uni Prot Number
FUNC	NM_002427.2	Human	P45452

Key Applications

Entrez Gene Number

Species

Uni Prot Number

FUNC

NM_002427.2

Human

P45452

Description
Catalogue Number	CC1047
Brand Family	Chemicon®
Trade Name	Chemicon
Description	MMP-13, human, proform, his-tagged, E. coli recombinant
Overview	CC1047 is a recombinant 452 amino acid polypeptide corresponding to human Pro-MMP-13, with an additional C-terminal His-tag with the sequence GVTHHHHHH expressed in E. coli and purified from periplasm. The calculated Mr of the recombinant protein is 51.681 kDa. Upon activation with APMA activated MMP-13 is formed.
Alternate Names	Procollagenase 3
Background Information	Matrix metalloproteinases (MMPs) are Zn2+- and Ca2+-dependent endopeptidases which function in the turnover of extracellular matrix components [Matrisian, 1992]. Main subfamilies of MMP are collagenases, gelatinases, stromelysins and membrane-type matrix metalloproteinases [Nagase, 1997]. Three homologous collagenases have been identified in human tissues: Interstitial collagenase, neutrophil collagenase and collagenase-3. These three enzymes cleave fibrillar collagens at a single site, generating fragments of approximately ¾ and ¼ the size of the original molecules. ProMMP-13 [Procollagenase-3] consists of 452 amino acids with a calculated Mr of 52.520 [Freije et al., 1994]. Due to N-linked glycosylation, the actual Mr is about 60000 Da [Knauper et al., 1996]. Within the protein the following domains and sequence regions can be distinguished [Freije et al., 1994; Knauper et al., 1996]: An N-terminal propeptide, which confers latency to the proenzyme, a Ca2+ and Zn2+-ion binding catalytic domain, a hinge region, and a C-terminal hemopexin-like domain. Latent procollagenase-3 can be activated by proteases such as stromelysin [ Knauper et al., 1996], gelatinase A, MT1-MMP and plasmin [ Knauper et al., 1996] or incubation with APMA Knauper et al., 1996]. The Mr of active collagenase-3 which begins with the N-terminal sequence YNVFPRTL is 48,000 Da. Collagenase-3 hydrolyzes type II collagen 5- to 6- times faster than type I and type III collagens. The enzyme also exhibits high activity towards gelatin and it degrades SERPINS as a1-antichymotrypsin and plasminogen activator inhibitor-2 [Knauper et al., 1996]. Collagenase-3 is inhibited in a 1:1 stoichiometric fashion by TIMP-1, TIMP-2 and TIMP-3. Collagenase-3 is expressed during fetal bone development [Stahlebackdahl, 1997]. In adult human tissues collagenase-3 has been detected only in pathological conditions: in malignant tumors [Freije et al., 1994], in chronic ulcers [Valaamo et al., 1997], in arthritic cartilage [Mitchell et al., 1996] and synovium [Wernicke et al., 1996].

Description

Catalogue Number

CC1047

Brand Family

Chemicon®

Trade Name

Chemicon

Description

MMP-13, human, proform, his-tagged, E. coli recombinant

Overview

CC1047 is a recombinant 452 amino acid polypeptide corresponding to human Pro-MMP-13, with an additional C-terminal His-tag with the sequence GVTHHHHHH expressed in E. coli and purified from periplasm. The calculated Mr of the recombinant protein is 51.681 kDa. Upon activation with APMA activated MMP-13 is formed.

Alternate Names

Procollagenase 3

Background Information

Matrix metalloproteinases (MMPs) are Zn2+- and Ca2+-dependent endopeptidases which function in the turnover of extracellular matrix components [Matrisian, 1992]. Main subfamilies of MMP are collagenases, gelatinases, stromelysins and membrane-type matrix metalloproteinases [Nagase, 1997]. Three homologous collagenases have been identified in human tissues: Interstitial collagenase, neutrophil collagenase and collagenase-3. These three enzymes cleave fibrillar collagens at a single site, generating fragments of approximately ¾ and ¼ the size of the original molecules.

ProMMP-13 [Procollagenase-3] consists of 452 amino acids with a calculated Mr of 52.520 [Freije et al., 1994]. Due to N-linked glycosylation, the actual Mr is about 60000 Da [Knauper et al., 1996]. Within the protein the following domains and sequence regions can be distinguished [Freije et al., 1994; Knauper et al., 1996]: An N-terminal propeptide, which confers latency to the proenzyme, a Ca2+ and Zn2+-ion binding catalytic domain, a hinge region, and a C-terminal hemopexin-like domain. Latent procollagenase-3 can be activated by proteases such as stromelysin [ Knauper et al., 1996], gelatinase A, MT1-MMP and plasmin [ Knauper et al., 1996] or incubation with APMA Knauper et al., 1996]. The Mr of active collagenase-3 which begins with the N-terminal sequence YNVFPRTL is 48,000 Da.

Collagenase-3 hydrolyzes type II collagen 5- to 6- times faster than type I and type III collagens. The enzyme also exhibits high activity towards gelatin and it degrades SERPINS as a1-antichymotrypsin and plasminogen activator inhibitor-2 [Knauper et al., 1996]. Collagenase-3 is inhibited in a 1:1 stoichiometric fashion by TIMP-1, TIMP-2 and TIMP-3.

Collagenase-3 is expressed during fetal bone development [Stahlebackdahl, 1997]. In adult human tissues collagenase-3 has been detected only in pathological conditions: in malignant tumors [Freije et al., 1994], in chronic ulcers [Valaamo et al., 1997], in arthritic cartilage [Mitchell et al., 1996] and synovium [Wernicke et al., 1996].

Product Information
Presentation	Provided as a liquid in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 0.05% Brij-35.

Product Information

Presentation

Provided as a liquid in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 0.05% Brij-35.

Applications
Key Applications	Affects Function
Application Notes	ACTIVATION: An aliquot of 19.5 μL procollagenase-3 is mixed with 0.5 μL APMA solution (40 mM p-aminophenyl mercuric acetate in DMSO) and the mixture is incubated for 30 minutes at 37°C. The mixture may be stored on ice until use for activity assays. INHIBITORS: MMP-13 is inhibited by TIMPs and by chelators of divalent cations such as EDTA or o-phenanthroline.

Applications

Key Applications

Affects Function

Application Notes

ACTIVATION:

An aliquot of 19.5 μL procollagenase-3 is mixed with 0.5 μL APMA solution (40 mM p-aminophenyl mercuric acetate in DMSO) and the mixture is incubated for 30 minutes at 37°C. The mixture may be stored on ice until use for activity assays.

INHIBITORS:

MMP-13 is inhibited by TIMPs and by chelators of divalent cations such as EDTA or o-phenanthroline.

Biological Information
Concentration	5 μg/25μL
Purity	Pro-MMP-13 appears as a major band at about 60 kDa in SDS-PAGE (>95% total protein).
Specific Activity	SPECIFIC ACTIVITY: <br /><br />The specific activity of activated MMP-13 is 250-300 mU/mg, where 1 U is the activity that hydrolyzes 1 μmol peptide (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-Dpa-Ala-Arg) within 1 minute (Knight, 1992).
Entrez Gene Number	NM_002427.2
Entrez Gene Summary	Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The protein encoded by this gene cleaves type II collagen more efficiently than types I and III. It may be involved in articular cartilage turnover and cartilage pathophysiology associated with osteoarthritis. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.
Gene Symbol	MMP13 MMP-13 CLG3 EC 3.4.24.-
UniProt Number	P45452
UniProt Summary	FUNCTION: SwissProt: P45452 # Degrades collagen type I. Does not act on gelatin or casein. Could have a role in tumoral process. COFACTOR: Binds 4 calcium ions per subunit. & Binds 2 zinc ions per subunit. SIZE: 471 amino acids; 53820 Da TISSUE SPECIFICITY: Seems to be specific to breast carcinomas. DOMAIN: SwissProt: P45452 The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme. DISEASE: SwissProt: P45452 # Defects in MMP13 are the cause of spondyloepimetaphyseal dysplasia type 2 (SEMD2) [MIM:602111]; also known as spondyloepimetaphyseal dysplasia type Missouri. SEMDs are a heterogeneous group of skeletal disorders characterized by defective growth and modeling of the spine and long bones. The SEMDs are distinguished from the spondylometaphyseal dysplasias and the spondyloepiphyseal dysplasias by the combined involvement of the epiphyses and metaphyses. The 3 disorders have malformations of the vertebrae in common. SIMILARITY: Belongs to the peptidase M10A family. & Contains 4 hemopexin-like domains.

Biological Information

Concentration

5 μg/25μL

Purity

Pro-MMP-13 appears as a major band at about 60 kDa in SDS-PAGE (>95% total protein).

Specific Activity

SPECIFIC ACTIVITY: <br /><br />The specific activity of activated MMP-13 is 250-300 mU/mg, where 1 U is the activity that hydrolyzes 1 μmol peptide (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-Dpa-Ala-Arg) within 1 minute (Knight, 1992).

Entrez Gene Number

NM_002427.2

Entrez Gene Summary

Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The protein encoded by this gene cleaves type II collagen more efficiently than types I and III. It may be involved in articular cartilage turnover and cartilage pathophysiology associated with osteoarthritis. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.

Gene Symbol

MMP13
MMP-13
CLG3
EC 3.4.24.-

UniProt Number

P45452

UniProt Summary

FUNCTION: SwissProt: P45452 # Degrades collagen type I. Does not act on gelatin or casein. Could have a role in tumoral process.
COFACTOR: Binds 4 calcium ions per subunit. & Binds 2 zinc ions per subunit.
SIZE: 471 amino acids; 53820 Da
TISSUE SPECIFICITY: Seems to be specific to breast carcinomas.
DOMAIN: SwissProt: P45452 The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
DISEASE: SwissProt: P45452 # Defects in MMP13 are the cause of spondyloepimetaphyseal dysplasia type 2 (SEMD2) [MIM:602111]; also known as spondyloepimetaphyseal dysplasia type Missouri. SEMDs are a heterogeneous group of skeletal disorders characterized by defective growth and modeling of the spine and long bones. The SEMDs are distinguished from the spondylometaphyseal dysplasias and the spondyloepiphyseal dysplasias by the combined involvement of the epiphyses and metaphyses. The 3 disorders have malformations of the vertebrae in common.
SIMILARITY: Belongs to the peptidase M10A family. & Contains 4 hemopexin-like domains.

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Maintain frozen at -70°C in undiluted aliquots. The enzyme may be stored at -20°C for several weeks without significant loss of activity. Repeated freezing and thawing should be avoided.

Storage and Shipping Information

Storage Conditions

Maintain frozen at -70°C in undiluted aliquots. The enzyme may be stored at -20°C for several weeks without significant loss of activity. Repeated freezing and thawing should be avoided.

Packaging Information
Material Size	5 µg

Packaging Information

Material Size

5 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

ProMMP-13 Recombinant Procollagenase-3

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