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Related Concept Videos

Phosphorylation01:02

Phosphorylation

51.4K
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
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Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

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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...
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Other Stress Responses in Bacteria01:30

Other Stress Responses in Bacteria

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Bacteria have global regulatory systems that control several types of stress mechanisms. These include Pho regulon and the heat shock response, which are essential systems for environmental adaptation, such as nutrient limitation and proteotoxic stress. The Pho regulon and the heat shock response exemplify bacterial resilience, enabling rapid adaptation to fluctuating environmental conditions.Pho RegulonBacteria require phosphorus for essential cellular processes, including nucleic acid...
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cAMP-dependent Protein Kinase Pathways01:25

cAMP-dependent Protein Kinase Pathways

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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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Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

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Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
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Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

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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...
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Related Experiment Video

Updated: Sep 27, 2025

A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues
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A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues

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Widespread Arginine Phosphorylation in Staphylococcus aureus.

Nadine Prust1, Pieter C van Breugel1, Simone Lemeer1

  • 1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands; Netherlands Proteomics Center, Utrecht, The Netherlands.

Molecular & Cellular Proteomics : MCP
|April 14, 2022
PubMed
Summary

Arginine phosphorylation, a key process in bacteria like Staphylococcus aureus, is now easier to study. Researchers identified over 1000 arginine phosphorylation sites, advancing our understanding of this widespread bacterial modification.

Keywords:
Fe(3+)-IMACLC-MS/MSStp1agrinine phosphorylationstaphylococcus aureus

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Arginine phosphorylation, a post-translational modification involving a phosphoramidate bond, plays a significant role in bacteria such as Bacillus subtilis and Staphylococcus aureus.
  • Large-scale analysis of arginine phosphorylation faces challenges due to substoichiometric levels, acid instability of phosphoramidates, and potential neutral losses during mass spectrometry.
  • Previous methods were hindered by the perceived instability of arginine phosphorylation, complicating enrichment and analysis.

Purpose of the Study:

  • To establish robust methods for the large-scale analysis of arginine phosphorylation in bacteria.
  • To identify and characterize arginine phosphorylation sites in methicillin-resistant Staphylococcus aureus (MRSA).
  • To investigate the role of the serine/threonine phosphatase Stp1 in regulating arginine phosphorylation.

Main Methods:

  • Utilized iron(III)-immobilized metal affinity chromatography (Fe3+-IMAC) for enrichment of phosphopeptides, demonstrating stability of arginine phosphorylation (pArg).
  • Employed higher-energy collisional dissociation (HCD) as the gold standard for analyzing phosphopeptides via LC-MS/MS.
  • Generated and used synthetic arginine phosphorylated peptides for validation of identified sites in S. aureus.

Main Results:

  • Demonstrated that arginine phosphorylation is stable enough for Fe3+-IMAC enrichment and LC-MS/MS analysis.
  • Identified 1062 arginine phosphorylation sites in S. aureus, creating the most comprehensive arginine phosphoproteome to date.
  • Validated the presence and localization of arginine phosphorylation in S. aureus using synthetic peptides.

Conclusions:

  • Arginine phosphorylation is a widespread and significant regulatory mechanism in bacteria, amenable to large-scale proteomic analysis.
  • The phosphatase Stp1 indirectly influences the arginine phosphoproteome in S. aureus, suggesting complex regulatory networks.
  • This study provides a foundation for further research into the functional roles of arginine phosphorylation in bacterial physiology and pathogenesis.