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

Hemoglobin01:24

Hemoglobin

7.4K
Hemoglobin is a globular protein made up of four subunits. Two of these subunits are alpha chains, and the other two are beta chains. Each subunit contains a molecule of heme, which has an iron atom and can bind to oxygen. When an oxygen molecule binds to one heme group, it changes the shape of hemoglobin, making it easier for the other heme groups to bind oxygen as well.
When all four heme groups are bound to oxygen, the resulting molecule is called oxyhemoglobin. As a result, arterial blood...
7.4K
Hematopoiesis01:21

Hematopoiesis

8.7K
The process of blood cell formation is called hematopoiesis. Hematopoiesis starts early during development, on the seventh day of embryogenesis. This phase of hematopoiesis is called the primitive wave, wherein the extraembryonic yolk sac allows the production of erythroid cells and endothelial cells from a common precursor called hemangioblast. The erythroid cells provide oxygen to support the growth of the rapidly dividing embryo. Hemangioblasts later develop into hematopoietic stem cells or...
8.7K
Colors and Magnetism03:02

Colors and Magnetism

13.9K
Color in Coordination Complexes
When atoms or molecules absorb light at the proper frequency, their electrons are excited to higher-energy orbitals. For many main group atoms and molecules, the absorbed photons are in the ultraviolet range of the electromagnetic spectrum, which cannot be detected by the human eye. For coordination compounds, the energy difference between the d orbitals often allows photons in the visible range to be absorbed and emitted, which is seen as colors by the human...
13.9K
Overview of Hematopoiesis01:20

Overview of Hematopoiesis

8.3K
Hematopoiesis, or blood cell production, is a vital biological process that begins early in embryonic development and continues throughout life. This process generates the various types of cells found in blood, including red blood cells, white blood cells, and platelets from hematopoietic stem cells (HSCs).
Developmental Phases of Hematopoiesis
Initially, HSCs are formed in the embryonic yolk sac, a critical site for early blood cell production. These stem cells subsequently migrate to other...
8.3K
Regulation of Hematopoietic Stem Cells01:01

Regulation of Hematopoietic Stem Cells

4.0K
All blood and immune cells are produced from the multipotent hematopoietic stem cells (HSCs) by the process of hematopoiesis. However, they all have a limited life span. In addition, many are depleted in immune surveillance or combatting an injury or infection. This makes blood one of the most regenerative tissues. Hematopoiesis helps replenish these blood and immune cells, restoring the body's normal functioning. However, overproduction of blood and immune cells can make them cancerous or...
4.0K
Multipotency of Hematopoietic Stem Cells01:19

Multipotency of Hematopoietic Stem Cells

3.8K
The hematopoietic stem cells or HSCs are multipotent, meaning they can differentiate and give rise to all blood and immune cells. HSCs are maintained in the quiescent stage until an external stimulus initiates their differentiation. The multipotent HSCs exist as two heterogeneous populations, long-term repopulating cells (LTRC) and short-term repopulating cells (STRC). The two HSC populations have different surface markers or receptors and are classified based on quiescence and long-term...
3.8K

You might also read

Related Articles

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

Sort by
Same author

Phage-assisted continuous evolution of enzymes for noncanonical tyrosine biosynthesis.

bioRxiv : the preprint server for biology·2026
Same author

Promiscuity-Guided Enzyme Evolution via Substrate Multiplexed Screening.

Angewandte Chemie (International ed. in English)·2026
Same author

Enzymatic Metal-Hydrogen Atom Transfer with a Cobalt Protoporphyrin Cofactor.

Journal of the American Chemical Society·2026
Same author

Enhanced Stereochemical Analysis of β‑Diastereomeric Amino Acids with Variants of Marfey's Reagent.

ACS omega·2025
Same author

Highly Sensitive Detection of Tyrosine and Neurotransmitters by Stereoselective Biosynthesis and Photochemically Induced Dynamic Nuclear Polarization.

Journal of the American Chemical Society·2025
Same author

Active site diversification of a non-canonical amino acid decarboxylase by merging substrate multiplexed screening with computationally guided recombination.

Protein science : a publication of the Protein Society·2025
Same journal

Clinical Europium fluorescent based lectin assays for mucin O-glycomics.

Methods in enzymology·2026
Same journal

A dual-color FRET assay for detection and quantitative analysis of O-glycopeptidases.

Methods in enzymology·2026
Same journal

Evolutionary genetic approaches to analyze mucins.

Methods in enzymology·2026
Same journal

Ex vivo imaging and enzymatic analysis of intestinal mucus.

Methods in enzymology·2026
Same journal

Glyco-TRAPP: A real-time glycocalyx permeability assay for assessing transmembrane mucin barrier function in live and fixed tissues.

Methods in enzymology·2026
Same journal

Quantitative imaging approaches to capture structural and functional dynamics of colonic mucus in health and disease in situ.

Methods in enzymology·2026
See all related articles

Related Experiment Video

Updated: Jan 16, 2026

Measurement of Heme Synthesis Levels in Mammalian Cells
09:43

Measurement of Heme Synthesis Levels in Mammalian Cells

Published on: July 9, 2015

12.5K

Cobalt-substituted hemoprotein expression.

Madeline E Rodemeier1, Olivia P Holsinger1, Andrew R Buller1

  • 1Department of Chemistry, University of Wisconsin-Madison, Madison, WI, United States.

Methods in Enzymology
|October 5, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed a method to replace iron with cobalt in hemoproteins, creating cobalt-substituted variants. This technique, using cobalt protoporphyrin IX (CoPPIX) in Escherichia coli, yields artificial metalloenzymes for advanced spectroscopic and structural studies.

Keywords:
Artificial MetalloenzymeCobalt ProtoporphyrinElectron Paramagnetic ResonanceHemeSynthetic Biology

More Related Videos

Analyzing Platelet Subpopulations by Multi-color Flow Cytometry
08:04

Analyzing Platelet Subpopulations by Multi-color Flow Cytometry

Published on: June 10, 2025

1.4K
Colony Forming Cell CFC Assay for Human Hematopoietic Cells
11:30

Colony Forming Cell CFC Assay for Human Hematopoietic Cells

Published on: December 18, 2010

36.3K

Related Experiment Videos

Last Updated: Jan 16, 2026

Measurement of Heme Synthesis Levels in Mammalian Cells
09:43

Measurement of Heme Synthesis Levels in Mammalian Cells

Published on: July 9, 2015

12.5K
Analyzing Platelet Subpopulations by Multi-color Flow Cytometry
08:04

Analyzing Platelet Subpopulations by Multi-color Flow Cytometry

Published on: June 10, 2025

1.4K
Colony Forming Cell CFC Assay for Human Hematopoietic Cells
11:30

Colony Forming Cell CFC Assay for Human Hematopoietic Cells

Published on: December 18, 2010

36.3K

Area of Science:

  • Biochemistry
  • Bioinorganic Chemistry

Background:

  • Metalloenzyme metal centers dictate reactivity and spectroscopic properties.
  • Cobalt substitution in hemoproteins offers complementary properties and novel reactivity.
  • Iron is the native metal in hemoproteins like myoglobin.

Purpose of the Study:

  • To establish a protocol for cobalt protoporphyrin IX (CoPPIX) biosynthesis and incorporation into hemoproteins.
  • To generate cobalt-substituted hemoproteins as artificial metalloenzymes.
  • To enable new spectroscopic and structural studies of hemoproteins.

Main Methods:

  • Biosynthesis and incorporation of CoPPIX into hemoproteins during expression in Escherichia coli BL21(DE3).
  • Utilized Physeter macrocephalus (sperm whale) myoglobin as a model hemoprotein.
  • Employed electronic absorption spectroscopy, LC-MS, ICP-MS, and EPR spectroscopy for analysis.

Main Results:

  • Successfully generated cobalt-substituted hemoproteins using the described protocol.
  • Demonstrated the effectiveness of the method for producing Co-substituted hemoproteins.
  • Validated analytical techniques for assessing cofactor identity and purity.

Conclusions:

  • The developed protocol provides an effective and accessible method for creating artificial metalloenzymes.
  • Cobalt-substituted hemoproteins are valuable tools for spectroscopic characterization and structural biology.
  • This method facilitates the production of useful quantities of Co-substituted hemoproteins.