Pro-MMP-9 Monomer

Key Applications	Entrez Gene Number	Species	Uni Prot Number
FUNC	NM_004994.2	Human	P14780

Key Applications

Entrez Gene Number

Species

Uni Prot Number

FUNC

NM_004994.2

Human

P14780

Description
Catalogue Number	CC1048
Brand Family	Chemicon®
Trade Name	Chemicon
Description	MMP-9, human, proform
Overview	CC1048 is isolated from human blood. The preparation is free from MMP-9 dimer and from complexes of MMP-9 with TIMP-1 or lipocalin. Upon activation with trypsin, enzymatically active MMP-9 of Mr 86 kDa is formed. INHIBITORS: MMP-9 is inhibited by TIMPs and by chelators of divalent cations, such as EDTA or o-phenanthroline.
Alternate Names	Progelatinase B 92 kDa Type IV Collagenase
Background Information	Human MMP-9 consists of 668 amino acids with a calculated Mrof 76 240 Da. Due to N- and O-linked glycosylated the Mr in SDS-PAGE is about 92 kDa [Wilhelm et al., 1989]. Within the protein sequence, the following structural domains can be distinguished [Wilhelm et al., 1989; Collier, et al., 1991]: An N-terminal propeptide, which confers latency to the proenzyme, a Ca2+ and Zn2+ ion-binding catalytic domain containing an insertion of three repeats homologous to type II repeats in the gelatin-binding region of fibronectin, and a C-terminal hemopexin-like domain. Catalytic and hemopexin domains are connected by a proline-rich sequence with homology to sequences in collagens. The gelatin-binding region and the hemopexin domain confer specific macromolecular substrate binding to MMP-9. The hemopexin domain of the latent enzyme binds TIMP-1 [Wilhelm et al., 1989; Goldberg et al., 1992; Kolkenbrock et al., 1995]. Activation of latent MMP-9 can be mediated by proteases like stromelysin, cathepsin G, kallikrein and trypsin or by incubation with APMA [Murphy & Grabbe, 1995]. In the presence of APMA, the propeptide is not removed completely, however, and there occurs considerable C-terminal self-processing [Murphy & Grabbe, 1995]. The enzyme is inhibited by TIMP-1 and TIMP-2. MMP-9 is secreted from macrophages, polymorphonuclear leukocytes, keratinocytes and many tumor cells [Hibbs et al., 1984; Sato et al., 1991; Sato et al., 1992]. It is detected in human plasma [Zucker et al., 1993] and saliva [Makela et al., 1994]. MMP-9 is involved in physiological processes such as angiogenesis, wound healing, bone remodeling, migration of macrophages and leukocytes [Shapiro, 1998]. Hydrolysis of type IV collagen and other matrix proteins in basement membranes by MMP-9 contributes to tumor cell invasion [Bernhard et al., 1994] and aortic aneurysm formation [Thompson, 1995].

Description

Catalogue Number

CC1048

Brand Family

Chemicon®

Trade Name

Chemicon

Description

MMP-9, human, proform

Overview

CC1048 is isolated from human blood. The preparation is free from MMP-9 dimer and from complexes of MMP-9 with TIMP-1 or lipocalin. Upon activation with trypsin, enzymatically active MMP-9 of Mr 86 kDa is formed.

INHIBITORS:

MMP-9 is inhibited by TIMPs and by chelators of divalent cations, such as EDTA or o-phenanthroline.

Alternate Names

Progelatinase B
92 kDa Type IV Collagenase

Background Information

Human MMP-9 consists of 668 amino acids with a calculated Mrof 76 240 Da. Due to N- and O-linked glycosylated the Mr in SDS-PAGE is about 92 kDa [Wilhelm et al., 1989]. Within the protein sequence, the following structural domains can be distinguished [Wilhelm et al., 1989; Collier, et al., 1991]: An N-terminal propeptide, which confers latency to the proenzyme, a Ca2+ and Zn2+ ion-binding catalytic domain containing an insertion of three repeats homologous to type II repeats in the gelatin-binding region of fibronectin, and a C-terminal hemopexin-like domain. Catalytic and hemopexin domains are connected by a proline-rich sequence with homology to sequences in collagens. The gelatin-binding region and the hemopexin domain confer specific macromolecular substrate binding to MMP-9. The hemopexin domain of the latent enzyme binds TIMP-1 [Wilhelm et al., 1989; Goldberg et al., 1992; Kolkenbrock et al., 1995].

Activation of latent MMP-9 can be mediated by proteases like stromelysin, cathepsin G, kallikrein and trypsin or by incubation with APMA [Murphy & Grabbe, 1995]. In the presence of APMA, the propeptide is not removed completely, however, and there occurs considerable C-terminal self-processing [Murphy & Grabbe, 1995]. The enzyme is inhibited by TIMP-1 and TIMP-2.

MMP-9 is secreted from macrophages, polymorphonuclear leukocytes, keratinocytes and many tumor cells [Hibbs et al., 1984; Sato et al., 1991; Sato et al., 1992]. It is detected in human plasma [Zucker et al., 1993] and saliva [Makela et al., 1994].

MMP-9 is involved in physiological processes such as angiogenesis, wound healing, bone remodeling, migration of macrophages and leukocytes [Shapiro, 1998]. Hydrolysis of type IV collagen and other matrix proteins in basement membranes by MMP-9 contributes to tumor cell invasion [Bernhard et al., 1994] and aortic aneurysm formation [Thompson, 1995].

Product Information
Presentation	Provided as a liquid in 50 mM Tris-HCl, pH 7.0, 200 mM NaCl, 5 mM CaCl2, ,1 μM ZnCl2, 0.05% Brij-35, 0.05% NaN3.

Product Information

Presentation

Provided as a liquid in 50 mM Tris-HCl, pH 7.0, 200 mM NaCl, 5 mM CaCl2, ,1 μM ZnCl2, 0.05% Brij-35, 0.05% NaN3.

Applications
Key Applications	Affects Function
Application Notes	ACTIVATION: An aliquot of 10 μL MMP-9 monomer is mixed with 20 μL trypsin solution (see below) and activation buffer in a total volume of 100 μL. The mixture is incubated for 20 min at 37°C. Thereafter trypsin is inhibited by addition of 10 μL aprotinin solution. Trypsin solution: 0.50 mg TPCK-trypsin/mL activation buffer. The solution is stored in aliquots at -20°C. Activation Buffer: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2. Aprotinin solution: 1 mg aprotinin/mL activation buffer. The solution is stored at -20°C.

Applications

Key Applications

Affects Function

Application Notes

ACTIVATION:

An aliquot of 10 μL MMP-9 monomer is mixed with 20 μL trypsin solution (see below) and activation buffer in a total volume of 100 μL. The mixture is incubated for 20 min at 37°C. Thereafter trypsin is inhibited by addition of 10 μL aprotinin solution.

Trypsin solution: 0.50 mg TPCK-trypsin/mL activation buffer. The solution is stored in aliquots at -20°C.

Activation Buffer: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2.

Aprotinin solution: 1 mg aprotinin/mL activation buffer. The solution is stored at -20°C.

Biological Information
Concentration	5 mg/25 mL
Purity	MMP-9 monomer appears as a major band at 92 kDa in non-reducing SDS-PAGE (>95% of total protein).
Specific Activity	SPECIFIC ACTIVITY: <br /><br />The specific activity of activated MMP-9 after trypsin activation is >1200 mU/mg, where 1 U is the activity that hydrolyzes 1 mmol peptide (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-Dpa-Ala-Arg) within 1 minute under the assay conditions described by Knight ,et al. When measured with the peptide substrate dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg [Masui et al., 1977] the specific activity of MMP-9 monomer is >1500 mU/mg.
Entrez Gene Number	NM_004994.2
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 enzyme encoded by this gene degrades type IV and V collagens. Studies in rhesus monkeys suggest that the enzyme is involved in IL-8-induced mobilization of hematopoietic progenitor cells from bone marrow, and murine studies suggest a role in tumor-associated tissue remodeling.
Gene Symbol	MMP9 MMP-9 GELB CLG4B EC 3.4.24.35 [Contains: 67 kDa matrix metalloproteinase-9 82 kDa matrix metalloproteinase-9].
UniProt Number	P14780
UniProt Summary	FUNCTION: SwissProt: P14780 # May play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1 at a Gly- -Leu bond. COFACTOR: Binds 2 zinc ions per subunit. & Binds 3 calcium ions per subunit. SIZE: 707 amino acids; 78427 Da SUBUNIT: Exists as monomer, disulfide-linked homodimer, and as a heterodimer with a 25 kDa protein. Macrophages and transformed cell lines produce only the monomeric form. TISSUE SPECIFICITY: Produced by normal alveolar macrophages and granulocytes. DOMAIN: SwissProt: P14780 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. PTM: Processing of the precursor yields different active forms of 64, 67 and 82 kDa. Sequentially processing by MMP3 yields the 82 kDa matrix metalloproteinase-9. SIMILARITY: Belongs to the peptidase M10A family. & Contains 3 fibronectin type-II domains. & Contains 4 hemopexin-like domains. MISCELLANEOUS: In the arthritis patient this enzyme might contribute to the pathogenesis of joint destruction and might constitute a useful marker of disease status.

Biological Information

Concentration

5 mg/25 mL

Purity

MMP-9 monomer appears as a major band at 92 kDa in non-reducing SDS-PAGE (>95% of total protein).

Specific Activity

SPECIFIC ACTIVITY: <br /><br />The specific activity of activated MMP-9 after trypsin activation is >1200 mU/mg, where 1 U is the activity that hydrolyzes 1 mmol peptide (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-Dpa-Ala-Arg) within 1 minute under the assay conditions described by Knight ,et al. When measured with the peptide substrate dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg [Masui et al., 1977] the specific activity of MMP-9 monomer is >1500 mU/mg.

Entrez Gene Number

NM_004994.2

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 enzyme encoded by this gene degrades type IV and V collagens. Studies in rhesus monkeys suggest that the enzyme is involved in IL-8-induced mobilization of hematopoietic progenitor cells from bone marrow, and murine studies suggest a role in tumor-associated tissue remodeling.

Gene Symbol

MMP9
MMP-9
GELB
CLG4B
EC 3.4.24.35 [Contains: 67 kDa matrix metalloproteinase-9
82 kDa matrix metalloproteinase-9].

UniProt Number

P14780

UniProt Summary

FUNCTION: SwissProt: P14780 # May play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1 at a Gly- -Leu bond.
COFACTOR: Binds 2 zinc ions per subunit. & Binds 3 calcium ions per subunit.
SIZE: 707 amino acids; 78427 Da
SUBUNIT: Exists as monomer, disulfide-linked homodimer, and as a heterodimer with a 25 kDa protein. Macrophages and transformed cell lines produce only the monomeric form.
TISSUE SPECIFICITY: Produced by normal alveolar macrophages and granulocytes.
DOMAIN: SwissProt: P14780 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.
PTM: Processing of the precursor yields different active forms of 64, 67 and 82 kDa. Sequentially processing by MMP3 yields the 82 kDa matrix metalloproteinase-9.
SIMILARITY: Belongs to the peptidase M10A family. & Contains 3 fibronectin type-II domains. & Contains 4 hemopexin-like domains.
MISCELLANEOUS: In the arthritis patient this enzyme might contribute to the pathogenesis of joint destruction and might constitute a useful marker of disease status.

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 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 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

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