Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

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

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...
Structural Protein Function01:56

Structural Protein Function

Structural proteins are a category of proteins responsible for functions ranging from cell shape and movement to providing support to major structures such as bones, cartilage, hair, and muscles. This group includes proteins such as collagen, actin, myosin, and keratin.
Collagen, the most abundant protein in mammals, is found throughout the body. In connective tissue, such as skin, ligaments, and tendons, it provides tensile strength and elasticity.  In bones and teeth, it mineralizes to form...
Structural Protein Function01:56

Structural Protein Function

Structural proteins are a category of proteins responsible for functions ranging from cell shape and movement to providing support to major structures such as bones, cartilage, hair, and muscles. This group includes proteins such as collagen, actin, myosin, and keratin.
Collagen, the most abundant protein in mammals, is found throughout the body. In connective tissue, such as skin, ligaments, and tendons, it provides tensile strength and elasticity.  In bones and teeth, it mineralizes to form...
Cytoskeletal Linker Proteins - Plakins01:09

Cytoskeletal Linker Proteins - Plakins

Plakins are large proteins with binding domains for microtubules, microfilaments, intermediate filaments, and membrane-associated protein complexes at cell junctions. Plakin functions are evolutionarily conserved and are primarily involved in organizing the different components of the cytoskeleton by crosslinking them to each other and connecting them to the cell-matrix and cell adhesion complexes. They are also known to interact with signal transducers, serve as scaffolds for signaling...
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Persistent and transient senescent cells contribute to brain-barrier development.

Cell·2026
Same author

A PKA-selective inhibitor captures an open but more ordered conformation of the PKA catalytic subunit.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

A catalogue of missense and nonsense mutation abundances for the U.S. cancer patient population.

medRxiv : the preprint server for health sciences·2026
Same author

Structural mechanism for noncanonical GPCR signaling in the Hedgehog pathway.

Nature structural & molecular biology·2026
Same author

Sex-specific disruptions in PKCγ signaling in a mouse model of spinocerebellar ataxia type 14.

JCI insight·2026
Same author

Cellular Prion Protein Engages the <i>N</i>-Methyl-d-Aspartate Receptor through N- and C-Terminal Domains.

Biochemistry·2026
Same journal

TDP-43 proteinopathy as a biomarker and therapeutic target in amyotrophic lateral sclerosis.

Biochemical Society transactions·2026
Same journal

Advancing the monitoring of organelle contact sites in vitro and in vivo.

Biochemical Society transactions·2026
Same journal

Mechanisms influencing transient cytoplasmic protein targeting to intracellular lipid droplets.

Biochemical Society transactions·2026
Same journal

Replication associated nuclear DNA mismatch repair across kingdoms.

Biochemical Society transactions·2026
Same journal

Phosphatases of regenerating liver downregulate PTEN to promote tumorigenesis.

Biochemical Society transactions·2026
Same journal

Implications of Rho GTPase signaling in cancer immunotherapy.

Biochemical Society transactions·2026
See all related articles

Related Experiment Video

Updated: May 9, 2026

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

Pseudokinases from a structural perspective.

Susan S Taylor1, Andrey Shaw, Jiancheng Hu

  • 1Department of Biomedical Sciences, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA. staylor@ucsd.edu

Biochemical Society Transactions
|July 19, 2013
PubMed
Summary
This summary is machine-generated.

Researchers engineered a kinase-dead B-Raf pseudokinase by mutating a conserved residue. This mutant retains scaffold function, demonstrating that kinase scaffold activity can drive downstream signaling independently of enzymatic activity.

More Related Videos

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O
03:09

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O

Published on: August 9, 2024

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
10:17

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

Related Experiment Videos

Last Updated: May 9, 2026

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O
03:09

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O

Published on: August 9, 2024

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
10:17

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • The catalytic subunit of PKA is a prototype for protein kinases.
  • Active kinases possess two hydrophobic spines: the regulatory spine (R-spine) and the catalytic spine (C-spine).
  • The C-spine is completed by the adenine ring of ATP.

Purpose of the Study:

  • To apply the 'spine' concept to engineer a kinase-dead B-Raf pseudokinase.
  • To investigate the role of B-Raf's catalytic versus scaffold functions in downstream signaling.
  • To explore a general strategy for creating pseudokinases.

Main Methods:

  • Mutation of a conserved C-spine residue (Ala481 to Phenylalanine) in B-Raf.
  • Assessment of ATP binding and kinase activity of the engineered mutant.
  • Analysis of B-Raf mutant dimerization and downstream signaling (MEK/ERK activation).

Main Results:

  • The A481F B-Raf mutant is kinase-dead, unable to bind ATP due to C-spine fusion.
  • The mutant retains the ability to bind wild-type B-Raf and C-Raf.
  • Dimerization with wild-type Raf activates downstream MEK and ERK signaling, independent of Ras and enzymatic activity.

Conclusions:

  • B-Raf is a bifunctional kinase with both catalytic and scaffold roles.
  • The scaffold function of B-Raf is sufficient for downstream signaling.
  • Conserved residue mutation offers a general strategy for engineering pseudokinases to study non-catalytic functions.