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Allosteric Proteins-ATCase

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Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...

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Lysosomes are the site for the degradation of macromolecules and biological polymers released during membrane trafficking events such as secretory, endocytic, autophagic, and phagocytic pathways. The membrane-enclosed area of the lysosome, called the lumen, contains hydrolytic enzymes active in an acidic environment. These acid hydrolases are functional at a pH between 4.5 and 5 and are involved in cellular processes such as cell signaling, energy metabolism, restoration of the plasma membrane,...

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An oxygen-based nucleophile, like water, can undergo addition reactions with aldehydes and ketones. The reaction leads to the formation of hydrates, also referred to as 1,1-diols or geminal diols.
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Allosteric Regulation

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Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...

Dehydration of Aldols to Enals: Base-Catalyzed Aldol Condensation

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This lesson delves into the aldol condensation catalyzed by bases, where aldols undergo dehydration to enals. As shown in Figure 1, the β-hydroxy aldehyde formed in a base-catalyzed aldol addition reaction dehydrates on heating to yield an unsaturated carbonyl product, which is commonly referred to as an enal.

Base-Catalyzed Aldol Addition Reaction

Base-Catalyzed Aldol Addition Reaction

As depicted in Figure 1, base-catalyzed aldol addition involves adding two carbonyl compounds in aqueous sodium hydroxide to form a β-hydroxy carbonyl compound.

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Related Experiment Video

Updated: May 10, 2026

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Published on: June 20, 2019

Structure and function of allophanate hydrolase.

Chen Fan¹, Zi Li¹, Huiyong Yin¹

¹From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.

The Journal of Biological Chemistry

|June 12, 2013

Summary

Allophanate hydrolase, crucial for urea metabolism and pathogen survival, has a newly elucidated crystal structure. This reveals its N and C domains

Keywords:

Allophanate Hydrolase Amidase Signature Family Decarboxylase Enzyme Catalysis Enzyme Structure Nitrogen Metabolism Urea Utilization X-ray Crystallography

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Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

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Published on: April 12, 2019

Area of Science:

Biochemistry
Structural Biology
Enzymology

Background:

Allophanate hydrolase is vital for organisms utilizing urea as a nitrogen source.
This enzyme plays roles in eukaryotic pyrimidine degradation, fungal dimorphism, and herbicide breakdown.
Understanding its structure and mechanism is key to diverse biological processes.

Purpose of the Study:

To determine the crystal structure of Kluyveromyces lactis allophanate hydrolase.
To elucidate the structure-function relationship and catalytic mechanism of the enzyme.
To gain insights into the enzyme's role in various biological pathways.

Main Methods:

X-ray crystallography to determine the enzyme's three-dimensional structure.
Structure-directed functional assays to probe enzyme activity and domain contributions.
Biochemical analysis to investigate the catalytic mechanism.

Main Results:

The crystal structure of Kluyveromyces lactis allophanate hydrolase was determined.
The N and C domains were shown to catalyze a two-step reaction and maintain enzyme dimeric form for optimal activity.
A novel decarboxylation reaction catalyzed by the C domain was identified.

Conclusions:

The dimeric structure of allophanate hydrolase, facilitated by its N and C domains, is essential for its function.
The study provides molecular insights into the enzyme's catalytic mechanism, including a novel decarboxylation reaction.
Findings expand the understanding of allophanate hydrolase's biological roles and catalytic capabilities.