recombinant human matriptase-2  (Enzo Biochem)

Journal: BMC Research Notes

Article Title: Matriptase cleaves the amyloid-beta peptide 1–42 at Arg-5, Lys-16, and Lys-28

doi: 10.1186/s13104-018-4040-z

Figure Lengend Snippet: Cleavage of Aβ1–42 by matriptase. a Synthetic human Aβ42 (1 µg, Bio-Techne) was mixed with a recombinant human matriptase protease domain (MatPD, “+” = 0.1 µg, “+++” = 0.3 µg, Bio-Techne) in a total volume of 20 µl with reaction buffer. Samples containing Aβ42 or MatPD alone were also prepared. The reactions were carried out at 37 °C for either overnight or 1.5 h, as indicated, before the samples were analyzed by SDS-PAGE and stained with Coomassie Brilliant Blue. b Duplicate samples as described for lanes 1 and 3 were sent for MALDI-TOF MS analysis, performed by the University of Florida Proteomics & Mass Spectrometry Core on a 4700 Proteomics Analyzer (Applied Biosystems). Peptide fragments generated by MatPD from Aβ42 were identified in the data graph from Sample 3 (Aβ42 + MatPD), as shown by the red boxes with the calculated (average) and observed masses listed in the table. Insets: left—separation of peaks “L17-K28” and “H6-K16” by MSight ; right—spectrum from Sample 1 (Aβ42 alone) set to the same intensity scale and in the same mass range. Other boxed peaks were also examined by MSight over the corresponding mass spectrum ranges between Samples 3 and 1 (not shown)

Article Snippet: Fig. 1 Cleavage of Aβ1–42 by matriptase. a Synthetic human Aβ42 (1 µg, Bio-Techne) was mixed with a recombinant human matriptase protease domain (MatPD, “+” = 0.1 µg, “+++” = 0.3 µg, Bio-Techne) in a total volume of 20 µl with reaction buffer.

Techniques: Recombinant, SDS Page, Staining, Mass Spectrometry, Generated

Journal: BMC Research Notes

Article Title: Matriptase cleaves the amyloid-beta peptide 1–42 at Arg-5, Lys-16, and Lys-28

doi: 10.1186/s13104-018-4040-z

Figure Lengend Snippet: Cleavage of APP in cultured cells by matriptase. a The APP695-EGFP (lanes 1, 3, 5, and 7) or the APP-R601A-EGFP mutant plasmid (R601A, lanes 2, 4, 6, and 8) (0.6 µg for each per transfection) was co-transfected in HEK293 cells with an empty plasmid (pLVX-Puro) (Vec, lanes 1 and 2), or the cDNA coding for human matriptase (Mat, lanes 3 and 4), protease-dead matriptase (MatM, lanes 5 and 6), or mouse TMPRSS6 (lanes 7 and 8) (0.4 µg for each per transfection). Twenty micrograms of total cell lysate from each sample were resolved on SDS-PAGE and western-blotted with an EGFP antibody or a β-tubulin antibody. The 55-kDa matriptase-specific APP-EGFP CTF is marked by the red asterisk, and the 38-kDa matriptase-specific APP-EGFP CTF is marked by the red arrowhead. b An empty plasmid (pLVX-Puro) (Vec, lane 1), or the cDNA coding for human matriptase (Mat, lane 2) or protease-dead matriptase (MatM, lane 3) (1.5 µg for each per transfection) was transfected in M17 human neuroblastoma cells. Twenty micrograms of total cell lysate from each sample were resolved on SDS-PAGE and western-blotted with an APP antibody, a matriptase antibody, or a GAPDH antibody. The data shown here were representative from three independently repeated experiments

Article Snippet: Fig. 1 Cleavage of Aβ1–42 by matriptase. a Synthetic human Aβ42 (1 µg, Bio-Techne) was mixed with a recombinant human matriptase protease domain (MatPD, “+” = 0.1 µg, “+++” = 0.3 µg, Bio-Techne) in a total volume of 20 µl with reaction buffer.

Techniques: Cell Culture, Mutagenesis, Plasmid Preparation, Transfection, SDS Page, Western Blot

Journal: Nature Communications

Article Title: SARS-CoV-2 infection of human lung epithelial cells induces TMPRSS-mediated acute fibrin deposition

doi: 10.1038/s41467-023-42140-6

Figure Lengend Snippet: A , B Expression of members of TMPRSS genes, ST14 and TMPRSS11D, in various cells as measured by A counts per 10 million total reads (TPM) from RNAseq, and B western blot. The expressions of TMPRSS genes in HEK293 and Vero cells are derived from GSE153744 and GSE83900 datasets from NCBI GEO databases. C pSARS-2 infection resulted in the release of soluble ST14 in the culture supernatant. D Metalloproteinase inhibitors, BB-94 and prinomastat, but not others inhibited infected NHBE cells induced fibrin clotting. The inhibitors were added during the viral infection but not during fibrin clotting assay. E Enzymatic cleavage of the prothrombin peptide, Thrb-324, by recombinant matriptase and HAT. F Recombinant matriptase and HAT cleaved prothrombin for fibrin clot formation similar to factor Xa. G Infection of 25 ng ST14 or 50 ng TMPRSS11D transfected ACE2-293T cells induced fibrin clot formation. Infected (I) or uninfected (UI) ACE2-293T cells without transfection did not form fibrin clots. All data are presented as mean values ± SD and unless stated all statistical analyses are performed using two-sided unpaired student t-test with p -values < 0.05 (*), <0.01 (**), <0.0001 (****).

Article Snippet: The cleavage of Thrb-324 peptide was initiated by mixing 10 μM of the peptide with 100 ng of human factor Xa (R&D Systems, Inc), or 400 ng of human matriptase (R&D Systems, Inc) in 100 μl assay buffer containing 25 mM Tris at pH 9.0, 2.5 μM ZnCl 2 , and 0.005% Brij-35 (w/v), or with infected cells or 100 μl of infected supernatant in 96-well plates.

Techniques: Expressing, Western Blot, Derivative Assay, Infection, Coagulation, Recombinant, Transfection

Journal: Scientific Reports

Article Title: Autophagy of Candida albicans cells after the action of earthworm Venetin-1 nanoparticle with protease inhibitor activity

doi: 10.1038/s41598-023-41281-4

Figure Lengend Snippet: Proteolytic activity of matriptase-1 ( A ) and matriptase-2 ( B ), trypsin ( C ), and chymotrypsin ( D ) after treatment with Venetin-1 versus the control sample without the inhibitor.

Article Snippet: Recombinant human matriptase-2 was obtained from Enzo Life Sciences (Switzerland).

Techniques: Activity Assay