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Related Concept Videos

Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes a mild...
cAMP-dependent Protein Kinase Pathways01:25

cAMP-dependent Protein Kinase Pathways

Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
Enzymes02:34

Enzymes

Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
Induced-fit Model01:13

Induced-fit Model

Most chemical reactions in cells require enzymes—biological catalysts that speed up the reaction without being consumed or permanently changed. They reduce the activation energy needed to convert the reactants into products. Enzymes are proteins, that usually work by binding to a substrate—a reactant molecule that they act upon.
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Enzyme Kinetics01:19

Enzyme Kinetics

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Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
11:11

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Published on: February 21, 2019

Creatine kinase structural changes induced by substrates.

Daniela Hornikova1, Petr Herman, Jiri Mejsnar

  • 1Faculty of Science, Charles University in Prague, Prague, Czech Republic.

Biochimica Et Biophysica Acta
|December 4, 2008
PubMed
Summary

Creatine kinase (CK) enzyme dynamics change with substrate binding, revealing three distinct conformations. These structural shifts are crucial for regulating cellular energy levels and enzyme activity.

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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes

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Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes

Published on: January 7, 2013

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Myofibrillar creatine kinase (CK) plays a vital role in buffering cellular adenosine triphosphate (ATP) concentrations during fluctuating muscle activity.
  • Understanding the structural dynamics of CK in response to substrate binding is essential for elucidating its regulatory mechanisms.

Purpose of the Study:

  • To investigate the structural and dynamic changes of the creatine kinase (CK) molecule upon binding with substrates ATP and creatine.
  • To identify distinct conformations of CK influenced by substrate ligation.

Main Methods:

  • Utilized steady-state and time-resolved fluorescence spectroscopy to examine the intrinsic tryptophan fluorescence of non-labelled CK.
  • Employed fluorescence quenching by acrylamide and anisotropy experiments with FITC-labelled CK to confirm structural dynamics.
  • Analyzed fluorescence lifetime and rotation correlation times to infer conformational states.

Main Results:

  • Observed significant differences in fluorescence lifetime and rotation correlation times for free CK, CK-ATP, and CK-ATP+creatine complexes.
  • Identified three distinct conformations of CK, with specific dynamic profiles for each substrate-bound state.
  • Confirmed substrate-dependent conformational changes using fluorescence quenching and anisotropy measurements.

Conclusions:

  • The study demonstrates that creatine kinase exists in at least three distinct conformations, dictated by the binding of ATP and creatine.
  • These conformational changes are proposed to follow an 'energy minimizing principle' driven by ligated substrates.
  • The findings reveal a critical aspect of subcellular physiological control, linking enzyme structure to reactivity.