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Related Concept Videos

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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Phosphorylation01:02

Phosphorylation

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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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Phosphoinositides and PIPs01:42

Phosphoinositides and PIPs

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Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...
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ATP Energy Storage and Release01:31

ATP Energy Storage and Release

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ATP is a highly unstable molecule. Unless quickly used to perform work, ATP spontaneously dissociates into ADP and inorganic phosphate (Pi), and the free energy released during this process is lost as heat. The energy released by ATP hydrolysis is used to perform work inside the cell and depends on a strategy called energy coupling. Cells couple the exergonic reaction of ATP hydrolysis with endergonic reactions, allowing them to proceed.
One example of energy coupling using ATP involves a...
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Riboswitches01:56

Riboswitches

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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
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Hydrolysis of ATP01:08

Hydrolysis of ATP

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The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids.
If one phosphate group is removed, a molecule of ADP—adenosine diphosphate—remains, along with inorganic phosphate. ADP can be further hydrolyzed to AMP—adenosine...
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Related Experiment Video

Updated: Aug 30, 2025

Chemical Triphosphorylation of Oligonucleotides
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Chemical Triphosphorylation of Oligonucleotides

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Polyphosphate Kinases Phosphorylate Thiamine Phosphates.

Jennie C Hildenbrand1, Georg A Sprenger1, Attila Teleki2

  • 1Institute of Microbiology, University of Stuttgart, Stuttgart, Germany.

Microbial Physiology
|August 30, 2022
PubMed
Summary
This summary is machine-generated.

Polyphosphate kinases (PPKs) can phosphorylate thiamine diphosphate (ThP2) and thiamine monophosphate (ThP1) into higher forms like thiamine triphosphate (ThP3). This reveals PPKs as promiscuous enzymes with potential roles in synthesizing various phosphorylated metabolites in vivo.

Keywords:
PolyphosphatePolyphosphate kinaseThiamine pyrophosphateThiamine triphosphateTransketolase

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Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry
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A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

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Area of Science:

  • Biochemistry
  • Enzymology

Background:

  • Polyphosphate kinases (PPKs) are enzymes involved in phosphate transfer reactions.
  • Their known function is to catalyze the transfer of the gamma-phosphate from nucleoside triphosphates to polyphosphate chains.

Purpose of the Study:

  • To investigate the substrate promiscuity of PPKs beyond their canonical function.
  • To explore the potential of PPKs in phosphorylating thiamine phosphates, such as thiamine diphosphate (ThP2) and thiamine monophosphate (ThP1).

Main Methods:

  • In vitro enzymatic assays using PPKs from various sources.
  • Phosphorylation of thiamine diphosphate (ThP2) and thiamine monophosphate (ThP1) using polyphosphate (polyP) as a phosphate donor.
  • High-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) for metabolite identification and mass determination.
  • Coupled enzyme assays to confirm the biological activity of synthesized thiamine metabolites.

Main Results:

  • PPKs were found to phosphorylate ThP2 to thiamine triphosphate (ThP3) and even thiamine tetraphosphate in vitro.
  • PPK2 enzymes, but not PPK1, efficiently phosphorylated ThP1 to ThP2 and ThP3.
  • The synthesized ThP2 demonstrated biological activity as a cofactor for the enzyme transketolase TktA.

Conclusions:

  • PPKs exhibit substrate promiscuity in vitro, extending their function to the phosphorylation of thiamine phosphates.
  • These findings suggest a potential in vivo role for PPKs in the biosynthesis of various phosphorylated thiamine metabolites.
  • The study highlights the versatility of PPKs and their potential involvement in novel metabolic pathways.