Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

Double mixing technique for linked enzyme assays.

R Landgraf¹, G F Betts

¹School of Biological Sciences, Queen Mary College, London, England.

Analytical Biochemistry

|February 1, 1989

Summary

A novel double mixing technique allows precise enzyme kinetics analysis by measuring product accumulation. This method enhances the study of enzyme interactions and product inhibition, particularly for 3-phosphoglycerate kinase.

Related Concept Videos

You might also read

Related Articles

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

Sort by

Same author

Enriched environment impacts trimethylthiazoline-induced anxiety-related behavior and immediate early gene expression: critical role of Crhr1.

The European journal of neuroscience·2014

Same author

Bidirectional rescue of extreme genetic predispositions to anxiety: impact of CRH receptor 1 as epigenetic plasticity gene in the amygdala.

Translational psychiatry·2014

Same author

Oxytocin in Brattleboro rats: increased synthesis is contrasted by blunted intrahypothalamic release from supraoptic nucleus neurones.

Journal of neuroendocrinology·2013

Same author

Plasma oxytocin and vasopressin do not predict neuropeptide concentrations in human cerebrospinal fluid.

Journal of neuroendocrinology·2013

Same author

Anxiety- rather than depression-like behavior is associated with adult neurogenesis in a female mouse model of higher trait anxiety- and comorbid depression-like behavior.

Translational psychiatry·2012

Same author

Truncated p110 ERBB2 induces mammary epithelial cell migration, invasion and orthotopic xenograft formation, and is associated with loss of phosphorylated STAT5.

Oncogene·2012

Area of Science:

Biochemistry
Enzymology
Chemical Kinetics

Background:

Enzyme kinetics are often studied using linked assays, which can introduce complexities.
Analyzing transient kinetics and enzyme interactions requires precise measurement of reaction rates.

Purpose of the Study:

To present a rapid double mixing technique for enzyme kinetics analysis.
To enable the determination of product accumulation rates during the delay between enzyme mixings.
To investigate enzyme interactions and product inhibition using this new method.

Main Methods:

Application of a rapid double mixing technique.
Enzymatic analysis of accumulated product after a delay.
Assay of 3-phosphoglycerate kinase kinetics.

Main Results:

The method allows determination of product accumulation rates during the mixing delay.
Steady-state and transient kinetic analyses are facilitated.
The assay of 3-phosphoglycerate kinase provided insights into its interaction with glyceraldehyde-3-phosphate dehydrogenase.

Conclusions:

The double mixing technique offers advantages for enzyme kinetics studies.
The results question proposed interactions between 3-phosphoglycerate kinase and glyceraldehyde-3-phosphate dehydrogenase.
The findings also raise questions regarding product inhibition in enzyme assays.

Related Experiment Videos