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Related Concept Videos

Calmodulin-dependent Signaling01:16

Calmodulin-dependent Signaling

Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
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...
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

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Protein kinases
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Activation and Inactivation of G Proteins01:22

Activation and Inactivation of G Proteins

Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high affinity and are together...
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...
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,...

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Related Experiment Video

Updated: May 21, 2026

Amide Hydrogen/Deuterium Exchange &amp; MALDI-TOF Mass Spectrometry Analysis of Pak2 Activation
07:15

Amide Hydrogen/Deuterium Exchange & MALDI-TOF Mass Spectrometry Analysis of Pak2 Activation

Published on: November 26, 2011

AMP-activated protein kinase undergoes nucleotide-dependent conformational changes.

Lei Chen1, Jue Wang, Yuan-Yuan Zhang

  • 1MOE Key Laboratory of Protein Sciences, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China.

Nature Structural & Molecular Biology
|June 5, 2012
PubMed
Summary

AMP-activated protein kinase (AMPK) is regulated by AMP binding to its gamma subunit. Nucleotide-binding site 3 is key for allosteric activation, with its occupancy differing between ATP and AMP.

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Amide Hydrogen/Deuterium Exchange &amp; MALDI-TOF Mass Spectrometry Analysis of Pak2 Activation
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Published on: November 26, 2011

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • AMP-activated protein kinase (AMPK) functions as a cellular energy sensor.
  • AMPK is a heterotrimeric complex regulated by AMP binding to the gamma subunit.

Purpose of the Study:

  • To investigate the structural basis of AMPK allosteric activation.
  • To determine the role of nucleotide-binding sites 3 and 4 in AMPK regulation.

Main Methods:

  • Cocrystallization of the mammalian AMPK core.
  • Mutagenesis studies of the gamma subunit.

Main Results:

  • Cocrystal structures revealed that nucleotide-binding site 3 on the gamma subunit is occluded in the presence of ATP.
  • Site 3 is occupied when AMP is bound to AMPK.
  • Mutagenesis data suggest sites 3 and 4 are crucial for allosteric activation.

Conclusions:

  • Nucleotide binding at site 3 is critical for AMPK allosteric activation.
  • The differential occupancy of site 3 by ATP versus AMP underlies AMPK's role as an energy sensor.