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The removal of time–concentration data points from progress curves improves the determination of K$_m$ : the example of paraoxonase 1
ID
Petrič, Boštjan
(
Avtor
),
ID
Goličnik, Marko
(
Avtor
),
ID
Bavec, Aljoša
(
Avtor
)
PDF - Predstavitvena datoteka,
prenos
(2,44 MB)
MD5: 5547C9460BC66F17F9DA9C2C38AFF4D7
URL - Izvorni URL, za dostop obiščite
https://www.mdpi.com/1420-3049/27/4/1306
Galerija slik
Izvleček
Several approaches for determining an enzyme’s kinetic parameter K$_m$ (Michaelis constant) from progress curves have been developed in recent decades. In the present article, we compare different approaches on a set of experimental measurements of lactonase activity of paraoxonase 1 (PON1): (1) a differential-equation-based Michaelis–Menten (MM) reaction model in the program Dynafit; (2) an integrated MM rate equation, based on an approximation of the Lambert W function, in the program GraphPad Prism; (3) various techniques based on initial rates; and (4) the novel program “iFIT”, based on a method that removes data points outside the area of maximum curvature from the progress curve, before analysis with the integrated MM rate equation. We concluded that the integrated MM rate equation alone does not determine kinetic parameters precisely enough; however, when coupled with a method that removes data points (e.g., iFIT), it is highly precise. The results of iFIT are comparable to the results of Dynafit and outperform those of the approach with initial rates or with fitting the entire progress curve in GraphPad Prism; however, iFIT is simpler to use and does not require inputting a reaction mechanism. Removing unnecessary points from progress curves and focusing on the area around the maximum curvature is highly advised for all researchers determining K$_m$ values from progress curves.
Jezik:
Angleški jezik
Ključne besede:
paraoxonase 1
,
dihydrocoumarin
,
progress curves
,
integrated Michaelis–Menten equation
,
Lambert W
,
lactonase activity
Vrsta gradiva:
Članek v reviji
Tipologija:
1.01 - Izvirni znanstveni članek
Organizacija:
MF - Medicinska fakulteta
Status publikacije:
Objavljeno
Različica publikacije:
Objavljena publikacija
Leto izida:
2022
Št. strani:
17 str.
Številčenje:
Vol. 27, iss. 4, art. 1306
PID:
20.500.12556/RUL-137296
UDK:
577
ISSN pri članku:
1420-3049
DOI:
10.3390/molecules27041306
COBISS.SI-ID:
98612227
Datum objave v RUL:
09.06.2022
Število ogledov:
731
Število prenosov:
96
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Objavi na:
Gradivo je del revije
Naslov:
Molecules
Skrajšan naslov:
Molecules
Založnik:
MDPI
ISSN:
1420-3049
COBISS.SI-ID:
18462981
Licence
Licenca:
CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:
http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:
To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.
Začetek licenciranja:
15.02.2022
Sekundarni jezik
Jezik:
Slovenski jezik
Ključne besede:
paraoksonaza 1
,
dihidrokumarin
,
krivulje napredka
Projekti
Financer:
ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Številka projekta:
P1-0170
Naslov:
Molekulski mehanizmi uravnavanja celičnih procesov v povezavi z nekaterimi boleznimi pri človeku
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