Selective Neurotensin-Derived Internally Quenched Fluorogenic Substrates for Neurolysin (EC 3.4.24.16): Comparison with Thimet Oligopeptidase (EC 3.4.24.15) and Neprilysin (EC 3.4.24.11)

2001 ◽  
Vol 292 (2) ◽  
pp. 257-265 ◽  
Author(s):  
Vitor Oliveira ◽  
Marcelo Campos ◽  
Jefferson P. Hemerly ◽  
Emer S. Ferro ◽  
Antonio C.M. Camargo ◽  
...  
Keyword(s):  
Fluorogenic Substrates ◽  
Thimet Oligopeptidase

scholarly journals Increased Stability of Oligopeptidases Immobilized on Gold Nanoparticles

Catalysts ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 78
Author(s):  
Marcelo Yudi Icimoto ◽  
Adrianne Marlise Mendes Brito ◽  
Marcos Paulo Cyrillo Ramos ◽  
Vitor Oliveira ◽  
Iseli Lourenço Nantes-Cardoso
Keyword(s):  
Gold Nanoparticles ◽  
Bioactive Peptides ◽  
Fluorogenic Substrates ◽  
Stabilizing Agents ◽  
Force Microscopy ◽  
Atomic Force ◽  
Plasmon Resonance Band ◽  
Resonance Band ◽  
Thimet Oligopeptidase

The metallopeptidases thimet oligopeptidase (THOP, EC 3.4.24.25) and neurolysin (NEL, EC 3.4.24.26) are enzymes that belong to the zinc endopeptidase M13 family. Numerous studies suggest that these peptidases participate in the processing of bioactive peptides such as angiotensins and bradykinin. Efforts have been conducted to develop biotechnological tools to make possible the use of both proteases to regulate blood pressure in mice, mainly limited by the low plasmatic stability of the enzymes. In the present study, it was investigated the use of nanotechnology as an efficient strategy for to circumvent the low stability of the proteases. Recombinant THOP and NEL were immobilized in gold nanoparticles (GNPs) synthesized in situ using HEPES and the enzymes as reducing and stabilizing agents. The formation of rTHOP-GNP and rNEL-GNP was characterized by the surface plasmon resonance band, zeta potential and atomic force microscopy. The gain of structural stability and activity of rTHOP and rNEL immobilized on GNPs was demonstrated by assays using fluorogenic substrates. The enzymes were also efficiently immobilized on GNPs fabricated with sodium borohydride. The efficient immobilization of the oligopeptidases in gold nanoparticles with gain of stability may facilitate the use of the enzymes in therapies related to pressure regulation and stroke, and as a tool for studying the physiological and pathological roles of both proteases.

Ultrasensitive Fluorogenic Substrates for Serine Proteases

1997 ◽  
Vol 78 (04) ◽  
pp. 1193-1201 ◽  
Author(s):  
Saulius Butenas ◽  
Maria E DiLorenzo ◽  
Kenneth G Mann
Keyword(s):  
Serine Proteases ◽  
Factor Viia ◽  
Activated Protein C ◽  
Factor Xa ◽  
High Specificity ◽  
High Rate ◽  
Fluorogenic Substrates ◽  
Type Plasminogen Activator ◽  
Factor Xia ◽  
Coagulation And Fibrinolysis

SummarySelective, sensitive assays for the quantitation of serine proteases involved in coagulation and fibrinolysis have been developed employing fluorogenic substrates containing a 6-amino-1-naphthalenesulfonamide leaving group (PNS-substrates). Over one hundred substrates were evaluated for hydrolysis by the serine proteases of blood coagulation and fibrinolysis, and substrate structure-efficiency correlations were examined. PNS-substrates which contain Lys in the P1 position are specific for Lys-plasmin and are either not hydrolyzed or hydrolyzed at a relatively low rate by factor Xa, thrombin, or urokinase-type plasminogen activator (uPA). These substrates allow quantitation of Lys-plasmin at concentrations as low as 1 pM. Eighteen of over 90 substrates tested for factor XIa are hydrolyzed by this enzyme at a relatively high rate reaching a kcat value of 170 s-1 and allowing quantitation of factor XIa at 10 fM. Eighteen of almost 90 PNS-substrates tested display high specificity for thrombin, some exceeding that for factor Xa by > 10,000-fold and > 100-fold for activated protein C (APC). Seven of these substrates have a over 100 s-1 and three of them have a KM below 1 μM. They allow the quantitation of thrombin at concentrations as low as 20 fM. For APC, uPA and the factor Vila/tissue factor complex, quantitation is feasible at 1 pM concentration. For factor Xa and factor VIIa the limits are 0.4 pM and 40 pM respectively. The PNS-substrates presented in this study may be employed for the development of direct and sensitive serine protease assays.

New fluorogenic substrates for renin

1981 ◽  
Vol 110 (1) ◽  
pp. 232-239 ◽  
Author(s):  
Kazuo Murakami ◽  
Tamiko Ohsawa ◽  
Shigehisa Hirose ◽  
Katsumi Takada ◽  
Shumpei Sakakibara
Keyword(s):  
Fluorogenic Substrates

An alternative approach for the synthesis of fluorogenic substrates of endo-β-(1→4)-xylanases and some applications

2008 ◽  
Vol 343 (3) ◽  
pp. 541-548 ◽  
Author(s):  
Mária Vršanská ◽  
Wim Nerinckx ◽  
Marc Claeyssens ◽  
Peter Biely
Keyword(s):  
Fluorogenic Substrates ◽  
Alternative Approach

Detecting enzymes in living cells using fluorogenic substrates

1993 ◽  
Vol 3 (3) ◽  
pp. 119-127 ◽  
Author(s):  
Richard P. Haugland ◽  
Iain D. Johnson
Keyword(s):  
Living Cells ◽  
Fluorogenic Substrates

4-Methylumbelliferyl esters as fluorogenic substrates for proteases

1983 ◽  
Vol 5 (2) ◽  
pp. 137-142 ◽  
Author(s):  
C.J. Gray ◽  
C.J.S.J. D'Silva ◽  
J. Boukouvalas ◽  
S.A. Barker
Keyword(s):  
Fluorogenic Substrates

Thimet Oligopeptidase and the Stability of MHC Class I Epitopes in Macrophage Cytosol

1999 ◽  
Vol 255 (3) ◽  
pp. 596-601 ◽  
Author(s):  
Fernanda C.V. Portaro ◽  
Marcelo D. Gomes ◽  
Adriana Cabrera ◽  
Beatriz L. Fernandes ◽  
Celio L. Silva ◽  
...  
Keyword(s):  
Mhc Class I ◽  
Class I ◽  
Thimet Oligopeptidase ◽  
The Stability
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