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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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IP3/DAG Signaling Pathway01:11

IP3/DAG Signaling Pathway

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Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and...
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Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

15.7K
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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Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

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Activation and Inactivation of G Proteins01:22

Activation and Inactivation of G Proteins

12.6K
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...
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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: Apr 15, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

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Two potential fish glycerol-3-phosphate phosphatases.

James A Raymond1

  • 1School of Life Sciences, University of Nevada Las Vegas, Las Vegas, NV, 89154, USA, raymond@unlv.nevada.edu.

Fish Physiology and Biochemistry
|April 3, 2015
PubMed
Summary

Researchers identified two glycerol-3-phosphate phosphatase (GPP) enzymes in winter-acclimated rainbow smelt. These enzymes may explain glycerol production for antifreeze but their activity at neutral pH is limited.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Zoology

Background:

  • Winter-acclimated rainbow smelt (Osmerus mordax) utilize glycerol as an antifreeze.
  • Glycerol production is typically facilitated by glycerol-3-phosphate phosphatase (GPP), an enzyme not previously identified in animals.

Purpose of the Study:

  • To identify and characterize glycerol-3-phosphate phosphatase (GPP) in rainbow smelt.
  • To investigate the potential role of identified GPP enzymes in glycerol-based antifreeze production in fish.

Main Methods:

  • Assembled and analyzed expressed sequence tag (EST) libraries from smelt.
  • Generated recombinant proteins for two candidate phosphatases.
  • Assayed enzyme activity and determined pH optima for recombinant proteins.

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Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
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Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
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Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation

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Main Results:

  • Identified two novel phosphatases in smelt with similarity to known GPP enzymes from algae and bacteria.
  • Recombinant proteins exhibited GPP activity, with acidic pH optima (~5.5).
  • Enzymes showed reduced activity at neutral pH, suggesting limited in vivo contribution to glycerol synthesis.

Conclusions:

  • Two GPP enzymes were identified in rainbow smelt, potentially explaining glycerol-based antifreeze.
  • The enzymes' low activity at neutral pH raises questions about their in vivo role in glycerol production.
  • Similar enzymes might be involved in glycerol production in other organisms, such as the insect Dendroctonus ponderosae.