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CC1048 Sigma-Aldrich

MMP-9, human, proform

Synonym: Progelatinase B, 92 kDa Type IV Collagenase

  •  eCl@ss 32160405

  •  NACRES NA.41

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Properties

Related Categories Application Index, Biochemicals and Reagents, Enzymes, Inhibitors, and Substrates, Protein & Nucleic Acid Isolation
Quality Level   100
biological source   human
assay   >95% (total protein)
specific activity   >1200 mU/mg (specific activity of activated MMP-9 after trypsin activation)
  >1500 mU/mg (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)
mfr. no.   Chemicon®
concentration   0.2 mg/mL
NCBI accession no.   NM_004994.2
UniProt accession no.   P14780
shipped in   dry ice
Gene Information   human ... MMP9(4318)

Description

General description

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.

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].

Application

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.

Unit Definition

Specific Activity: 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

Physical form

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.

Storage and Stability

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.

Analysis Note

MMP-9 monomer appears as a major band at 92 kDa in non-reducing SDS-PAGE

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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.

Safety & Documentation

Safety Information

WGK Germany 
WGK 1
Protocols & Articles
Peer-Reviewed Papers
15

References

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