Related Experiment Video

Updated: Aug 25, 2026

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting
10:08

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Published on: December 9, 2022

Regulation of NDR2 protein kinase by multi-site phosphorylation and the S100B calcium-binding protein

Mario R Stegert1, Rastislav Tamaskovic, Samuel J Bichsel

  • 1Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH 4058 Basel, Switzerland.

The Journal of Biological Chemistry
|March 24, 2004
PubMed

Insights

Nuclear Dbf2-related (NDR) protein kinases, including the newly characterized NDR2, regulate cell division. NDR2 activation involves specific phosphorylation events and is stimulated by S100B, similar to NDR1.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Nuclear Dbf2-related (NDR) protein kinases are AGC group kinases crucial for cell division and morphology.
  • NDR1 and NDR2 are highly conserved mammalian isoforms with distinct expression patterns.

Purpose of the Study:

  • To clone and characterize the human and mouse NDR2 protein kinase.
  • To investigate the activation mechanisms and regulation of NDR2.

Main Methods:

  • Cloning and sequence analysis of human and mouse NDR2.
  • Expression pattern analysis in mouse tissues.
  • Activation studies using okadaic acid and site-directed mutagenesis.
  • In vitro kinase assays with S100B.
  • Subcellular localization studies.

Main Results:

  • NDR1 and NDR2 share 86% amino acid identity but exhibit different tissue expression.
  • NDR2 activation by okadaic acid involves phosphorylation at Ser-282 (autophosphorylation) and Thr-442 (upstream kinase).
  • A constitutively active NDR2 mutant was generated by hydrophobic motif replacement.
  • S100B stimulated NDR2 autophosphorylation in vitro, indicating shared regulatory mechanisms with NDR1.
  • Ectopically expressed NDR2 showed predominant cytoplasmic localization.

Conclusions:

  • NDR2 is a critical regulator of cell division with distinct expression and activation pathways.
  • Phosphorylation of Ser-282 and Thr-442 are key events in NDR2 activation.
  • NDR2 regulation is similar to NDR1, involving S100B and specific phosphorylation sites.

More Related Videos

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag08:55

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Published on: December 14, 2017

Oligopeptide Competition Assay for Phosphorylation Site Determination09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

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,...
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
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...
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...
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

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.
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a rapamycin-insensitive companion...