The determination of proteolytic activity of snake venom by means of a chromogenic substrate

Toxicon ◽  
1973 ◽  
Vol 11 (1) ◽  
pp. 103-105 ◽  
Author(s):  
K. Steyn ◽  
G.R. Delpierre
Keyword(s):  
Proteolytic Activity ◽  
Snake Venom ◽  
Chromogenic Substrate

scholarly journals A PROCEDURE FOR THE DETERMINATION OF PROTEOLYTIC ACTIVITY

1950 ◽  
Vol 182 (2) ◽  
pp. 821-828
Author(s):  
Beat M. Iselin ◽  
Carl. Niemann
Keyword(s):  
Proteolytic Activity

One-pot hydrazide-based native chemical ligation for efficient chemical synthesis and structure determination of toxin Mambalgin-1

2014 ◽  
Vol 50 (44) ◽  
pp. 5837-5839 ◽  
Author(s):  
Man Pan ◽  
Yao He ◽  
Ming Wen ◽  
Fangming Wu ◽  
Demeng Sun ◽  
...  
Keyword(s):  
Chemical Synthesis ◽  
Snake Venom ◽  
Structure Determination ◽  
Native Chemical Ligation ◽  
One Pot ◽  
Chemical Ligation ◽  
Venom Toxin ◽  
Snake Venom Toxin ◽  
Switch Strategy

An efficient one-pot chemical synthesis of snake venom toxin Mambalgin-1 was achieved using an azide-switch strategy combined with hydrazide-based native chemical ligation.

scholarly journals Determination of digestive proteolytic profile in the larvae of Dyspessa palidata (Staudinger) (Lepidoptera: Cossidae)

2015 ◽  
Vol 47 (3) ◽  
pp. 91
Author(s):  
M. Mardani-Talaee ◽  
A. Zibaee
Keyword(s):  
Proteolytic Activity ◽  
Cathepsin B ◽  
Posterior Midgut ◽  
Ph Dependency ◽  
Metalloproteinase Inhibitors ◽  
Optimal Value ◽  
Important Pest ◽  
Proteolytic Activities ◽  
Membrane Bound

Digestive proteolytic profile was determined in the larvae of <em>Dyspessa palidata</em> (Staudinger), which is the most important pest of Alliaceae in Europe and Iran. Compartmentalisation of the proteolytic activities by considering soluble and membrane-bound fractions revealed that soluble fractions of the whole midgut preparations had higher general proteolytic activity than membrane-bound fractions. Also, four proteolytic bands were observed in the soluble fraction of the total midgut preparation in electrophoresis. Compartmentalisation of the specific proteases revealed presence of trypsin, elastase, aminoand carboxy peptidases in posterior midgut but the highest activities of other proteases were found in anterior midgut. The highest activity of general protease was found at pHs of 6 and 8. Also, pH dependency of trypsin, chymotrypsin and elastase were found at values of 8, 7-8 and 9 but cathepsins had the optimal pH at 6. Exopeptidases showed the optimal value at pH of 7 although carboxypeptidase showed same activity at values of 6 and 7. The inhibitory concentrations 50% (IC<sub>50</sub>) of AEBSF.HCL on trypsin, chymotrypsin and elastase proteases were found to be 3.69, 3.31 and 4.09 mM, respectively. IC<sub>50</sub> concentrations of TLCK, SBTI and TPCK significantly inhibited trypsin and chymotrypsin activities. IC<sub>50</sub> of E-64 were 3.67 and 4.16 mM on cathepsin B and L but cystatin revealed 5.22 and 4.48 mM concentrations on cathepsin B and L, respectively. EDTA and phenathroline as metalloproteinase inhibitors had IC<sub>50</sub> of 3.25 and 3.91 mM on general proteolytic activity. Results of the current study revealed larvae of <em>D. palidata</em> utilised different proteases to increase digestive efficiency when they fed on the host plants containing several toxic molecules.

scholarly journals Amplification of Color Yield Obtained Using a Chromogenic Substrate for Determining Plasminogen Activator

1977 ◽  
Author(s):  
Robert B. Friedmaa ◽  
Hau C. Kwaan ◽  
Marija Szczecinski
Keyword(s):  
Absorption Maximum ◽  
Enzyme Assay ◽  
Amide Bond ◽  
Chromogenic Substrate ◽  
Present Communication ◽  
Red Color ◽  
Enzyme Digest ◽  
Color Yield ◽  
Ammonium Sulfamate

A synthetic chromogenic substrate has recently been reported for use in the specific measurement of urokinase. This material (Chromozym UK, marketed by Pentapharm, Ltd., Basle) is a synthetic tripeptide linked through an amide bond to the chromogen para-nitroaniline (PNA). The release of free PNA can be monitored spectrophotometrically by measuring the increase of absorbance of light in the range of 380 to 410 nm. The present communication describes our efforts at enhancing the color yield of the assay as well as changing the absorption maximum to a wavelength which can more readily and accurately be measured on inexpensive laboratory spectrophotometers .Bratton and Marshall have developed a method which can be used for the determination of aromatic amines in biological systems. We have modified this method for use in semimicro applications and have adapted it to the measurement of free PNA in solution. After acidification of an aliquot of the enzyme digest with trichloroacetic acid (TCA), the liberated PNA is diazotized with sodium nitrite. Excess nitrous acid is removed with ammonium sulfamate, and the diazotized PNA is reacted with 1-naphthylethylenediamine reagent. The product has a magenta-red color with an absorption maximum of 545 nm. The substrate is not affected by exposure to TCA, and when fresh reagents are used, a low background is obtained. The color yield during actual enzyme assay is amplified twenty fold. Other coupling agents are also currently under investigation.

Determination of alpha 2-macroglobulin-trypsin complex with a new synthetic substrate.

1989 ◽  
Vol 35 (11) ◽  
pp. 2169-2172 ◽  
Author(s):  
K Kuroiwa ◽  
S Nakatsuyama ◽  
K Katayama ◽  
T Nagasawa
Keyword(s):  
Healthy Subjects ◽  
Colorimetric Assay ◽  
Colorimetric Method ◽  
Pancreatic Disease ◽  
Chromogenic Substrate ◽  
Standard Curve ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Abstract We have developed a colorimetric assay for quantifying alpha 2-macroglobulin-trypsin complex (alpha 2M-TRY) in human serum, based on use of a new chromogenic substrate D-gamma-tert-butyloxy-Glu-Gly-Arg-3-carboxy-4-hydroxyanilide dihydrochloride (PS-3001). Within-run CVs by this assay were 4.76%, 1.57%, and 0.83% for trypsin complex concentrations of 3.1, 12.2, and 48.1 U/L, respectively (n = 10 each). Between-day CVs were 5.38%, 3.12%, and 2.20% at each concentration, respectively (n = 7). Mean analytical recoveries of alpha 2M-TRY added to serum were 100%, 105%, and 101% for 9.2, 15.1, and 46.3 U/L, respectively (n = 2). The standard curve obtained was linear up to 330 U/L. We applied this method to the study of alpha 2M-TRY activity in sera from 97 healthy subjects (group 1), from 27 patients with acute pancreatitis (group 2), and from 25 patients with other chylopoietic diseases (group 3); results ranged from 0 to 1.2 U/L (mean = 0.5, SD = 0.3), from 1.2 to 77.4 U/L (mean = 14.6, SD = 19.0), and from 0 to 1.3 U/L (mean = 0.4, SD = 0.3), respectively. Concentrations of enzymatically active alpha 2M-TRY were significantly greater in sera from group 2 than in groups 1 and 3. The determination of serum alpha 2M-TRY activity by this simple, rapid, colorimetric method may be useful for the diagnosis and evaluation of pancreatic disease.

scholarly journals Proteolytic Activity Matrix Analysis (PrAMA) for simultaneous determination of multiple protease activities

2011 ◽  
Vol 3 (4) ◽  
pp. 422-438 ◽  
Author(s):  
Miles A. Miller ◽  
Layla Barkal ◽  
Karen Jeng ◽  
Andreas Herrlich ◽  
Marcia Moss ◽  
...  
Keyword(s):  
Simultaneous Determination ◽  
Proteolytic Activity ◽  
Matrix Analysis ◽  
Activity Matrix

A study on the degradation of arachin by proteolytic enzymes

1973 ◽  
Vol 51 (11) ◽  
pp. 2217-2222 ◽  
Author(s):  
R. B. van Huystee
Keyword(s):  
Amino Acids ◽  
Proteolytic Activity ◽  
Free Amino Acids ◽  
Free Amino ◽  
Proteolytic Enzymes ◽  
Disc Electrophoresis ◽  
Polyacrylamide Gels ◽  
Macromolecular Protein

The prime purpose of this proteolysis study was to direct attention to alternate means of measuring proteolytic activity other than the determination of free amino acids. The release of peptides from a macromolecular protein during incubation with either papain, pronase, or trypsin was determined by measuring the presence of 280-nm-absorbing molecules in the fractionation range of Sephadex G 25 eluant after incubation. The formation of larger proteinaceous constituents by proteolysis of arachin was analyzed by disc electrophoresis on polyacrylamide gels. Using these techniques it was noted that papain was the most efficient proteolytic agent for the degradation of arachin.

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