Detecting Hyaluronidase and Hyaluronidase Inhibitors: Hyaluronan-Substrate Gel and -Inverse Substrate Gel Techniques

2003 ◽  
pp. 391-397 ◽  
Author(s):  
Kazuhiro Mio ◽  
Antonei B Csóka ◽  
Susan Stair Nawy ◽  
Robert Stern
Keyword(s):  
Inverse Substrate

scholarly journals An active-site titrant for human tissue-type plasminogen activator

1986 ◽  
Vol 239 (2) ◽  
pp. 477-479 ◽  
Author(s):  
R A Smith
Keyword(s):  
Plasminogen Activator ◽  
Active Site ◽  
Human Tissue ◽  
Tissue Type ◽  
Tissue Type Plasminogen Activator ◽  
Rapid Release ◽  
Type Plasminogen Activator ◽  
Inverse Substrate

The reaction of recombinant tissue-type plasminogen activator with the inverse substrate 4-amidino-2-nitrophenyl 4′-anisate results in the rapid release of the chromogen 4-amidino-2-nitrophenol and the accumulation of the relatively stable 4-anisoyl-enzyme. Spectrophotometric monitoring of the reaction enables the operational molarity of the enzyme to be determined.

scholarly journals In Vitro Selection of Cathepsin E-Activity-Enhancing Peptide Aptamers at Neutral pH

2011 ◽  
Vol 2011 ◽  
pp. 1-10 ◽  
Author(s):  
Madhu Biyani ◽  
Masae Futakami ◽  
Koichiro Kitamura ◽  
Tomoyo Kawakubo ◽  
Miho Suzuki ◽  
...  
Keyword(s):  
In Vitro Selection ◽  
Physiological Activity ◽  
Peptide Aptamers ◽  
Neutral Ph ◽  
Cathepsin E ◽  
Analysis System ◽  
Ph Range ◽  
Cancerous Cells ◽  
Inverse Substrate

The aspartic protease cathepsin E has been shown to induce apoptosis in cancer cells under physiological conditions. Therefore, cathepsin E-activity-enhancing peptides functioning in the physiological pH range are valuable potential cancer therapeutic candidates. Here, we have used a general in vitro selection method (evolutionary rapid panning analysis system (eRAPANSY)), based on inverse substrate-function link (SF-link) selection to successfully identify cathepsin E-activity-enhancing peptide aptamers at neutral pH. A successive enrichment of peptide activators was attained in the course of selection. One such peptide activated cathepsin E up to 260%, had a high affinity (KD; ∼300 nM), and had physiological activity as demonstrated by its apoptosis-inducing reaction in cancerous cells. This method is expected to be widely applicable for the identification of protease-activity-enhancing peptide aptamers.

scholarly journals The Mechanism of Mus81-Mms4 Cleavage Site Selection Distinguishes It from the Homologous Endonuclease Rad1-Rad10

2003 ◽  
Vol 23 (10) ◽  
pp. 3487-3496 ◽  
Author(s):  
Suzanne A. Bastin-Shanower ◽  
William M. Fricke ◽  
Janet R. Mullen ◽  
Steven J. Brill
Keyword(s):  
Cleavage Site ◽  
Replication Fork ◽  
Synthetic Lethality ◽  
Excision Repair ◽  
Nucleotide Excision ◽  
Homologous Structure ◽  
Dna Strand ◽  
Mutant Phenotypes ◽  
Inverse Substrate

ABSTRACT Mus81-Mms4 and Rad1-Rad10 are homologous structure-specific endonucleases that cleave 3′ branches from distinct substrates and are required for replication fork stability and nucleotide excision repair, respectively, in the yeast Saccharomyces cerevisiae. We explored the basis of this biochemical and genetic specificity. The Mus81-Mms4 cleavage site, a nick 5 nucleotides (nt) 5′ of the flap, is determined not by the branch point, like Rad1-Rad10, but by the 5′ end of the DNA strand at the flap junction. As a result, the endonucleases show inverse substrate specificity; substrates lacking a 5′ end within 4 nt of the flap are cleaved poorly by Mus81-Mms4 but are cleaved well by Rad1-10. Genetically, we show that both mus81 and sgs1 mutants are sensitive to camptothecin-induced DNA damage. Further, mus81 sgs1 synthetic lethality requires homologous recombination, as does suppression of mutant phenotypes by RusA expression. These data are most easily explained by a model in which the in vivo substrate of Mus81-Mms4 and Sgs1-Top3 is a 3′ flap recombination intermediate downstream of replication fork collapse.

scholarly journals An Inverse Substrate Orientation for the Regioselective Acylation of 3′,5′-Diaminonucleosides Catalyzed by Candida antarctica lipase B?

ChemBioChem ◽  
2005 ◽  
Vol 6 (8) ◽  
pp. 1381-1390 ◽  
Author(s):  
Iván Lavandera ◽  
Susana Fernández ◽  
Julia Magdalena ◽  
Miguel Ferrero ◽  
Romas J. Kazlauskas ◽  
...  
Keyword(s):  
Candida Antarctica ◽  
Candida Antarctica Lipase B ◽  
Candida Antarctica Lipase ◽  
Lipase B ◽  
Regioselective Acylation ◽  
Substrate Orientation ◽  
Inverse Substrate

Modeling of the velocity profile of a bioreactor: the concept of biochemical process

2019 ◽  
Author(s):  
Chem Int
Keyword(s):  
Velocity Profile ◽  
Substrate Concentration ◽  
Discharge Coefficient ◽  
Linear Trend ◽  
Biomass Concentration ◽  
Space Velocity ◽  
Lag Phase ◽  
Biochemical Process ◽  
Model Equations ◽  
Inverse Substrate

This research covers the modeling of the velocity profile of a bioreactor with recycle; the concept of biochemical process. The biochemical process adopted is fermentation and a plug-flow fermenter (PFF) was taken as a case study. The derivation of workable model equations for monitoring and predicting the velocity profile of a PFF were obtained, together with obtaining the model equations for investigating the effect of microbial and substrate concentrations on the discharge coefficient, bioreactor’s volume. Constant data were sourced from literatures, together with hypothetical values to simulate the derived model equations using Mathlab. The substrate concentration decreases with time as biomass population increases with time. Effect of biomass concentration on discharge coefficient, shows that increase in biomass concentration brings a corresponding increase in the discharge coefficient as well as the bioreactor’s volume revealed that substrate concentration is depleting alongside with bioreactor’s volume follows the same trend of change when substrate concentration is decreasing irrespective of whether the length or area of the bioreactor is varied. The effect of microbial concentration on bioreactor volume when area and length of bioreactor are varied reveals that the process followed same trend only that there is a presence of lag phase upon the influence of inhibitors. The inverse substrate concentration increases, the space velocity also increases, and there is a linear trend of change on the inverse substrate concentration with respect to space velocity.

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