In Vitro Metabolism
In Vitro Metabolism - Related Videos
10:44
Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures
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2017
Metabolic competency of in vitro systems is a key requirement for the biotransformation and disposition of drugs and toxicants. In this protocol, we describe the application of reference metabolic probes to assess phase I metabolism in cell cultures.
05:41
Metabolic Profile Analysis of Zebrafish Embryos
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Cited by 42 •
2013
Zebrafish represent a powerful vertebrate model that has been under-utilised for metabolic studies. Here we describe a rapid way to measure the in vivo metabolic profile of developing zebrafish that allows the comparison of different mitochondrial function parameters between genetically or pharmacologically manipulated embryos, thereby increasing the applicability of this organism.
00:52
What is Metabolism?
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2019
Overview Metabolism represents all of the chemical activity in a cell, including reactions that build molecules (anabolism) and those that break molecules down (catabolism). Anabolic reactions require energy, whereas catabolic reactions provide it. Thus, metabolism describes how cells transform energy through a variety of chemical reactions, which are often made more efficient with the help of enzymes. Metabolism Is the Sum of All of the Chemical Reactions That Take Place in an Organism
10:19
An Introduction to Cell Metabolism
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2023
In cells, critical molecules are either built by joining together individual units like amino acids or nucleotides, or broken down into smaller components. Respectively, the reactions responsible for this are referred to as anabolic and catabolic. These reactions require or produce energy typically in the form of a “high-energy” molecule called ATP. Together, these processes make up “Cell Metabolism,” and are hallmarks of healthy, living cells.JoVE’s introduction to cell metabolism briefly...
08:28
Metabolic Labeling
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2023
Metabolic labeling is used to probe the biochemical transformations and modifications that occur in a cell. This is accomplished by using chemical analogs that mimic the structure of natural biomolecules. Cells utilize analogs in their endogenous biochemical processes, producing compounds that are labeled. The label allows for the incorporation of detection and affinity tags, which can then be used to elucidate metabolic pathways using other biochemical analytical techniques, such as SDS-PAGE...
FAQs
The enzyme system determines which substrate transformations occur and which metabolites accumulate. Oxidation, reduction, hydrolysis, and conjugation represent different reaction routes, so the enzyme components present can shape the resulting product profile. Examining these products helps biochemists connect enzyme activity with particular metabolic pathways and assess whether a molecule undergoes formation, modification, or clearance under controlled conditions.
These conditions influence both reaction rates and the products detected. Cofactors support enzyme-dependent transformations, while pH and temperature affect the environment in which the reactions proceed. Incubation time determines how long substrates remain available for conversion. Controlling these variables allows researchers to compare enzyme activity consistently and distinguish changes in metabolism from changes caused by assay conditions.
Isolated enzymes, subcellular fractions, cultured cells, and tissue preparations provide different levels of biological context. An isolated enzyme can clarify a specific activity, whereas a cell or tissue preparation can reflect the combined behavior of multiple metabolic components. Selecting among these systems helps match the experiment to its purpose, such as studying one reaction or examining broader metabolite formation.
These reaction categories indicate the chemical routes available for transforming a substrate. Identifying which route occurs, and which products result, can reveal how a molecule moves through a metabolic pathway. In biochemistry, this information supports characterization of enzyme activity and helps explain whether the tested system favors particular forms of molecular modification or clearance.
A typical design begins by selecting an appropriate enzyme, subcellular fraction, cultured cell, or tissue preparation and introducing the substrate. Researchers then establish suitable cofactors, pH, temperature, and incubation time before examining the metabolites formed or cleared. Keeping these factors defined creates a controlled comparison for evaluating reaction rates, products, and pathway behavior.
Drug discovery programs use these assays to predict how candidate compounds may be biotransformed and to identify metabolites that could contribute to toxicity. The results can also show how metabolism differs between species, helping researchers interpret experimental findings and refine compound structures. Together, these outcomes support the design of compounds with improved safety and effectiveness.