Name: graphing enzyme kinetics data
Uploaded: 2023-04-30T10:09:42.000Z
Duration: 1 min 19 s
Channel: JoVE

K12 Standards

north-carolina-k-12-science-standard-course-of-study

North Carolina K-12 Science Standard Course of Study

9-12

Biology

from-molecules-to-organisms-structures-and-processes

From Molecules to Organisms- Structures and Processes

LS.Bio.1.2

Introduction to Enzyme Kinetics

Enzim kinetiği, enzim katalizli reaksiyonların oranlarını inceler. Değişen substrat konsantrasyonlarında tekrarlanan deneylerin oranları, tüketilen substratın veya zaman içinde oluşan ürünün konsantrasyonu ölçülerek izlenir.
Bu sonuçlar, substrat konsantrasyonunun bir reaksiyonun hızını veya hızını nasıl etkilediğini göstermek için grafiklendirilebilir.
Reaksiyon hızı, düşük konsantrasyonlarda artan substrat miktarları ile doğrusal olarak artar, ancak daha yüksek konsantrasyonlarda plato yapmaya başlar. Hız, maksimum hıza veya Vmaks'a yaklaşır - enzimin substrat ile tamamen doyduğu hız.
Enzim afinitesi, bir enzimin substratını ne kadar güçlü veya zayıf bir şekilde bağladığını ölçer ve Michaelis sabiti olan KM ile ölçülür. KM değeri, oran Vmax'ın %50'si olduğunda substrat konsantrasyonuna eşittir.
Küçük bir KM, bir enzimin yüksek bir substrat afinitesine sahip olduğunu gösterir ve bunun tersi de geçerlidir. Daha büyük bir KM'ye sahip bir enzim, daha düşük bir K M'ye sahip bir enzime kıyasla maksimum hızına yaklaşmak için daha yüksek substrat konsantrasyonları gerektirir.

<ol>
	<li>Voet D. ve Voet J.G. Biyokimya, 4. baskı John Wiley & Sons, 2010 s-487-492</li>
	<li>Berg J.M., Tymoczko J.L. ve Stryer L. Biyokimya, 5. baskı, W.H. Freeman and Co., Bölüm 8.4</li>
</ol>

enzim kinetiğine giriş

Enzim kinetiği biyokimyasal reaksiyonların hızlarını inceler. Bilim adamları, çeşitli substrat konsantrasyonlarında belirli bir enzimatik reaksiyonun reaksiyon hızlarını izler. Reaksiyon hızını etkileyen inhibitörler veya diğer moleküllerle ilave denemeler de yapılabilir. Deneyci daha sonra reaksiyon özelliklerinin bir grafiğini elde etmek için belirli bir denemenin başlangıç reaksiyon hızını veya hızını (Vo) substrat konsantrasyonuna ([S]) göre çizebilir. Tek bir substratı içeren birçok enzimatik reaksiyon için bu veriler, Leonor Michaelis ve Maud Menten tarafından türetilen bir denklem olan Michaelis-Menten denklemine uyar. <img alt="Eq1" src="/files/ftp_upload/12164/12164_Equation 1.png" style="text-align: center; height: 70px;"> Denklem, incelenen enzim için maksimum hızı (Vmax) ve Michaelis sabitini (KM) tahmin eder ve aşağıdaki varsayımlara dayanır: <ol> <li>Reaksiyonun başlangıcında ürün mevcut değildir.</li> <li>Enzim-substrat kompleksi oluşum hızı, ayrışma ve ürünlere parçalanma hızına eşittir.</li> <li>Enzim konsantrasyonu, substrat konsantrasyonuna kıyasla minimum düzeydedir.</li> <li>Yalnızca başlangıç reaksiyon hızları ölçülür.</li> <li>Enzim ya serbest formda ya da enzim-substrat kompleksinde bulunur.</li> </ol> Lineweaver-Burke, Eadie-Hofsteot ve Hanes-Woolf grafikleri gibi Michaelis-Menten denkleminin farklı yeniden düzenlemeleri, kinetik parametrelerin grafiğini çizmenin alternatif yollarıdır. Lineweaver-Burke veya çift karşılıklı grafik genellikle KM ve Vmax'ı tahmin etmek için kullanılır. Grafik, Michaelis-Menten grafiğindeki x ve y ekseninin karşılıklı değerlerini kullanır. Matematiksel olarak y-kesme noktası 1/Vmax'a, x-kesme noktası ise −1/KM'ye eşittir. Lineweaver-Burke grafiği, yarışmalı, sınırlı yarışmalı ve yarışmasız inhibasyon türleri arasında görsel olarak ayrım yapmak için kullanılabilir. Kinetik parametreleri belirlemek için Michaelis-Menten denkleminin Eadie-Hofstee ve Hanes-Woolf grafikleri gibi farklı yeniden düzenlemeleri de kullanılır.

Enzim Kinetiğine Giriş

Enzim kinetiği, biyokimyasal reaksiyonların hızlarını inceler. Bilim adamları, çeşitli substratlarda belirli bir enzimatik reaksiyon için reaksiyon hızlarını izler ...

Education

JoVE Core

Cell Biology

Hücre Biyolojisi

Unit 1

Ünite 1

Energy and Catalysis

Enerji ve Kataliz

KEY TERMS AND CONCEPTS

TEMEL TERİMLER VE KAVRAMLAR

graphing enzyme kinetics data

Enzyme kinetics uses graphs to show how fast a biochemical reaction happens. Scientists measure the initial reaction rate, or velocity (Vo), at different substrate concentrations ([S]). They may also run trials with inhibitors or other molecules that change the reaction rate.

The results are often plotted as Vo against [S]. For many single-substrate reactions, the data fits the Michaelis-Menten equation. This equation, named after Leonor Michaelis and Maud Menten, helps estimate the maximum velocity (Vmax) and the Michaelis constant (KM) for the enzyme being studied.

The Michaelis-Menten model is based on a few assumptions. No product is present at the start of the reaction. The rate of enzyme-substrate complex formation equals the rate of its breakdown into products and its dissociation. The enzyme concentration is minimal compared with the substrate concentration. The model also uses only the initial reaction rates. In this setup, the enzyme is either free or bound in an enzyme-substrate complex.

Scientists can also use other graph forms to analyze the same data. The Lineweaver-Burke plot, also called the double reciprocal plot, uses the reciprocal values from the Michaelis-Menten graph. On this plot, the y-intercept equals 1/Vmax and the x-intercept equals -1/KM. It is often used to estimate KM and Vmax and to visually distinguish competitive, non-competitive, and uncompetitive inhibitors.

Other rearrangements of the Michaelis-Menten equation include the Eadie-Hofstee plot and the Hanes-Woolf plot. These graphs are also used to determine kinetic parameters. Together, these methods help scientists compare reaction behavior and understand how enzymes respond to substrate concentration and inhibitors.

Graphing Enzyme Kinetics Data

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key terms and concepts

Learn how enzyme kinetics data is graphed with Michaelis-Menten, Lineweaver-Burke, Eadie-Hofstee, and Hanes-Woolf plots to estimate Vmax and KM.

Graphing Enzyme Kinetics Data | JoVE K12

cell biology

JoVE Core Hücre Biyolojisi, anlaşılması kolay ancak kapsamlı animasyonlu video derslerde karmaşık hücresel süreçleri gösterir. Bu koleksiyon, organeller ve zarlar, hücresel taşıma ve hücre enerjileri dahil olmak üzere hücrelerin temel bileşenlerine ve işlevlerine odaklanır. Gerçek dünya araştırma deneylerinde hücre biyolojisinin uygulanmasını gösteren bilim insanı eylem videoları ile tamamlanmaktadır.

Virginia  Standards of Learning

HIgh School

BIO.2

BIO.2.c

Chemistry

CH.7

CH.7.f

Florida Department of Education

High School

Life Science

SC.912.L.18

SC.912.L.18.4

SC.912.L.18.11

Texas Essential Knowledge and Skills

BIO.11

BIO.11.B

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Graphing Enzyme Kinetics Data

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