MMP-1 蛋白, Human (HEK293, His)

The MMP-1 protein acts as an enzyme capable of cleaving types I, II, and III collagen at specific sites within the helical domain. In addition, it exhibits lytic activity against type VII and type X collagen. MMP-1 Protein, Human (HEK293, His) is the recombinant human-derived MMP-1 protein, expressed by HEK293 , with C-6*His labeled tag.

基质金属蛋白酶-1 (MMP-1) 是一种蛋白水解酶，因其在细胞外基质重塑中的作用而闻名。它表现出广泛的底物特异性，在螺旋域内的特定位点切割 I、II 和 III 型胶原蛋白。此外，MMP-1 表现出针对 VII 型和 X 型胶原蛋白的活性。值得注意的是，在 HIV 感染的情况下，MMP-1 与分泌的病毒 Tat 蛋白相互作用并裂解它，从而减少神经元 Tat 介导的神经毒性。这种相互作用强调了 MMP-1 除了其在细胞外基质降解中的典型作用之外的多种功能，表明它参与了与病毒感染和神经保护相关的过程（参考：未提供具体参考）。

Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2. Read at excitation and emission wavelengths of 360 nm and 405 nm. The specific activity is 4879.964 pmol/min/µg, as measured under the described conditions. (Activation description: The proenzyme needs to be activated by APMA (HY-148905) for an activated form.)

Materials
Assay buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB) Mercury p-aminophenyl acetate (APMA) , solubilized to 100 mM with DMSO
Substrate:MOCAc-PLGL(Dpa)AR (HY-131498)
Standard:7-amino, 4-Methyl Coumarin (AMC, HY-D0027)

Experimental steps
1. Standard curve: Dilute standard to 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, 100 μmol/L with assay buffer, and add 100 μL of each into black zymography wells. The excitation and emission wavelengths were 380 nm and 460 nm, respectively, and read in kinetic mode for 5 min. The measured RFU was used as the vertical coordinate, and the amount of standard material was used as the horizontal coordinate to make a standard curve and obtain the standard curve formula.
2. Human MMP-1 was diluted to 50 μg/mL with experimental buffer containing 1 mM of APMA.
3. Activation: incubate at 37°C for 2 hours.
4. Dilute the activated Human MMP-1 protein to 1ng/μL with buffer.
5. Dilute the substrate to 20 μM with experimental buffer.
6. Experimental group: 50 μL, 1 ng/μL Human MMP-1 protein + 50μL, 20 μM of substrate.
Control group: 50μL, 20 μM of substrate + 50μL of buffer.
7. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm, respectively.
8. Calculate the specific activity:

Human

HEK293

C-6*His

P03956 (F20-N469)

4312  [NCBI]

Full Length of Mature Protein

FPATLETQEQDVDLVQKYLEKYYNLKNDGRQVEKRRNSGPVVEKLKQMQEFFGLKVTGKPDAETLKVMKQPRCGVPDVAQFVLTEGNPRWEQTHLTYRIENYTPDLPRADVDHAIEKAFQLWSNVTPLTFTKVSEGQADIMISFVRGDHRDNSPFDGPGGNLAHAFQPGPGIGGDAHFDEDERWTNNFREYNLHRVAAHELGHSLGLSHSTDIGALMYPSYTFSGDVQLAQDDIDGIQAIYGRSQNPVQPIGPQTPKACDSKLTFDAITTIRGEVMFFKDRFYMRTNPFYPEVELNFISVFWPQLPNGLEAAYEFADRDEVRFFKGNKYWAVQGQNVLHGYPKDIYSSFGFPRTVKHIDAALSEENTGKTYFFVANKYWRYDEYKRSMDPGYPKMIAHDFPGIGHKVDAVFMKDGFFYFFHGTRQYKFDPKTKRILTLQKANSWFNCRKN

49-61 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

溶液

Supplied as a 0.2 μm filtered solution of 20 mM MES, 150 mM NaCl, 2 mM CaCl₂, 1 mM DTT, 0.05% Brij35, 10% Glycerol, pH 5.0 or 20 mM MES, 150 mM NaCl, 2 mM CaCl₂, 1 mM DTT, 10% Glycerol, pH 7.4.

<1 EU/μg, determined by LAL method.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.