GO:0016747 acyltransferase activity, transferring groups other than amino-acyl groups: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0016747 describes a molecular function: the catalysis of acyl group transfer from a donor to an acceptor, excluding amino-acyl transfer.
• This activity is essential for diverse metabolic and biosynthetic pathways, including natural product biosynthesis and tRNA modification.
• Enzymes with this activity often exhibit broad substrate tolerance, enabling them to process various acyl donors and acceptors.
• Studying GO:0016747 requires integrating biochemical assays, structural biology, and CRISPR-based genetic screens.
• Dysregulation of acyltransferases is linked to metabolic disorders and cancer, making them potential therapeutic targets.
• EDITGENE provides CRISPR knockout, point mutation, knock-in, overexpression, and library screening services to dissect GO:0016747-related gene functions.
Description
Acyltransferase activity, transferring groups other than amino-acyl groups (GO:0016747) is a fundamental molecular function that catalyzes the transfer of an acyl group from a donor molecule to an acceptor, excluding the transfer of amino-acyl groups. This activity is distinct from aminoacyl-tRNA synthetases and is involved in a wide range of biological processes, from secondary metabolite biosynthesis to post-translational modifications. Understanding this function is critical for researchers studying metabolic engineering, enzyme evolution, and drug discovery. The QuickGO definition provides a precise scope: catalysis of the transfer of an acyl group, other than amino-acyl, from one compound (donor) to another (acceptor). This definition encompasses enzymes such as acetyltransferases, acyltransferases, and related transferases that do not act on aminoacyl substrates. In this article, we explore the mechanism, key genes, disease relevance, and research methods for GO:0016747, with a focus on how CRISPR-based models can accelerate discovery.
acyltransferase activity, transferring groups other than amino-acyl groups At A Glance
| GO ID | GO:0016747 |
|---|---|
| GO term | acyltransferase activity, transferring groups other than amino-acyl groups |
| Ontology | molecular_function |
| Synonym | transferase activity, transferring acyl groups other than amino-acyl groups; transferase activity, transferring groups other than amino-acyl groups |
| Major function | Catalysis of acyl group transfer from donor to acceptor, excluding amino-acyl transfer |
| Definition | Catalysis of the transfer of an acyl group, other than amino-acyl, from one compound (donor) to another (acceptor). |
| Related enzymes | Acetyltransferases, acyltransferases, and other transferases not acting on aminoacyl substrates |
| Pathways | Natural product biosynthesis, lipid metabolism, tRNA modification, and secondary metabolism |
What Is GO:0016747?
GO:0016747 is a molecular function term defined as the catalysis of the transfer of an acyl group, other than amino-acyl, from one compound (donor) to another (acceptor). In simpler terms, it describes enzymes that move acyl groups (such as acetyl, malonyl, or other acyl chains) between molecules, but specifically excludes the transfer of amino acids to tRNA or other acceptors. This activity is central to many biosynthetic pathways, including the production of antibiotics, lipids, and modified proteins.
Why Is acyltransferase activity, transferring groups other than amino-acyl groups Important in Cell Biology?
GO:0016747 is important because it governs a vast array of biochemical reactions that are essential for cellular metabolism, natural product biosynthesis, and post-translational modifications. Enzymes with this activity are often involved in the production of bioactive molecules, including antibiotics and signaling lipids, and their dysregulation can lead to metabolic diseases and cancer. Moreover, understanding the substrate specificity and catalytic mechanism of these enzymes is crucial for drug design and metabolic engineering.
• Enables the biosynthesis of diverse natural products, including antibiotics and secondary metabolites.
• Plays a key role in tRNA modification and translational fidelity.
• Involved in lipid metabolism and membrane homeostasis.
• Contributes to post-translational modifications that regulate protein function.
• Dysregulation is associated with metabolic disorders and cancer.
• Provides targets for antimicrobial and anticancer drug development.
• Facilitates metabolic engineering for industrial biotechnology.
• Offers a model system for studying enzyme evolution and substrate promiscuity.
What Happens During acyltransferase activity, transferring groups other than amino-acyl groups?
Substrate Binding and Donor Selection
In simple terms: The enzyme grabs the acyl group donor and the acceptor molecule.
The first step in acyltransferase activity is the binding of the acyl donor and the acceptor substrate. For example, in dehydrophos biosynthesis, the enzyme binds an acyl donor such as acetyl-CoA and a specific acceptor, positioning them for catalysis. In Escherichia coli ItaT, the enzyme specifically recognizes aminoacyl-tRNAs as acceptors, but the acyl donor is not an aminoacyl group, distinguishing it from aminoacyl-tRNA synthetases. The binding pocket often accommodates a range of acyl donors, contributing to substrate promiscuity.
Catalytic Transfer of the Acyl Group
In simple terms: The enzyme moves the acyl group from the donor to the acceptor.
Once bound, the enzyme catalyzes the transfer of the acyl group from the donor to the acceptor. This typically involves a nucleophilic attack by the acceptor on the acyl donor, forming a new ester or amide bond. In the case of ItaT, the enzyme acetylates aminoacyl-tRNAs, transferring an acetyl group to the amino acid moiety. The reaction mechanism may involve a catalytic triad or a metal ion, depending on the enzyme family.
Product Release and Enzyme Turnover
In simple terms: The modified acceptor is released, and the enzyme is ready for another round.
After the acyl group is transferred, the modified acceptor is released from the active site, and the enzyme returns to its initial state to catalyze another reaction. In dehydrophos biosynthesis, the acylated product is further processed by downstream enzymes. The efficiency of product release can influence the overall catalytic rate and is often regulated by conformational changes.
Substrate Specificity and Proofreading
In simple terms: The enzyme checks that it has the right molecules before and after the reaction.
Many acyltransferases exhibit proofreading mechanisms to ensure fidelity. For instance, ItaT discriminates between different aminoacyl-tRNAs, acetylating only specific ones. This specificity is achieved through interactions between the enzyme and the acceptor molecule, often involving conserved residues in the active site. Understanding these specificity determinants is key for engineering enzymes with tailored substrate ranges.
Key Genes Involved in GO:0016747 acyltransferase activity, transferring groups other than amino-acyl groups
The following genes and proteins are representative examples of enzymes with acyltransferase activity, transferring groups other than amino-acyl groups, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ItaT (E. coli) | Acetylates aminoacyl-tRNAs, involved in tRNA modification | Model for substrate specificity and tRNA acetylation |
| Dehydrophos biosynthetic enzymes | Catalyze amide bond formation during dehydrophos biosynthesis | Studying natural product biosynthesis and acyl transfer |
| Acetyl-CoA acetyltransferase | Transfers acetyl groups in lipid metabolism | Target for metabolic engineering and drug discovery |
| Malonyl-CoA acyltransferase | Involved in fatty acid biosynthesis | Potential target for antibiotics |
| Chloramphenicol acetyltransferase | Inactivates chloramphenicol by acetylation | Antibiotic resistance marker |
| Gentamicin acetyltransferase | Acetylates aminoglycosides | Antibiotic resistance mechanism |
| Serine acetyltransferase | Catalyzes cysteine biosynthesis | Plant and microbial metabolism |
| Homoserine acetyltransferase | Methionine biosynthesis | Microbial metabolic engineering |
| Carnitine acyltransferase | Fatty acid oxidation | Metabolic disorders |
| Platelet-activating factor acetyltransferase | Lipid signaling | Inflammation |
| Histone acetyltransferase | Chromatin modification | Gene regulation and cancer |
| N-acetyltransferase | Drug metabolism | Pharmacogenetics |
| Aminoglycoside acetyltransferase | Antibiotic resistance | Clinical microbiology |
| Acyltransferase from Mycobacterium tuberculosis | Mycolic acid biosynthesis | Tuberculosis drug target |
| Fatty acyl-CoA synthetase | Activates fatty acids for acyl transfer | Lipid metabolism |
| Lysophospholipid acyltransferase | Membrane remodeling | Inflammation and cancer |
| Acyl-CoA:cholesterol acyltransferase | Cholesterol esterification | Atherosclerosis |
| Diacylglycerol acyltransferase | Triglyceride synthesis | Obesity and diabetes |
How Is acyltransferase activity, transferring groups other than amino-acyl groups Regulated?
The activity of enzymes with GO:0016747 is regulated at multiple levels, including transcriptional control, post-translational modifications, and allosteric regulation by metabolites. For example, the expression of ItaT in E. coli is induced under specific growth conditions, and its activity is modulated by the availability of aminoacyl-tRNAs. In dehydrophos biosynthesis, the acyltransferase activity is tightly coordinated with other biosynthetic enzymes to ensure efficient production of the final product. Additionally, feedback inhibition by downstream products and covalent modification by phosphorylation are common regulatory mechanisms.
acyltransferase activity, transferring groups other than amino-acyl groups and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| Histone acetyltransferase (HAT) | Leukemia, solid tumors | Knockout in cancer cell lines, xenograft models |
| Diacylglycerol acyltransferase (DGAT) | Obesity, insulin resistance | Knockout mice, overexpression in adipocytes |
| Carnitine acyltransferase | Cardiomyopathy, metabolic myopathy | Point mutation knock-in mice, patient-derived iPSCs |
| Aminoglycoside acetyltransferase | Antibiotic resistance | Knockout in bacteria, reporter assays |
| Acyl-CoA:cholesterol acyltransferase (ACAT) | Atherosclerosis | Knockout mice, overexpression in macrophages |
Acyltransferases in Cancer
Dysregulation of acyltransferase activity has been implicated in various cancers. For instance, histone acetyltransferases (HATs) are often mutated or aberrantly expressed in leukemia and solid tumors, leading to altered gene expression. Similarly, acyltransferases involved in lipid metabolism, such as diacylglycerol acyltransferase (DGAT), contribute to cancer cell proliferation and survival. Targeting these enzymes with small molecule inhibitors is an active area of anticancer drug discovery.
Acyltransferases in Metabolic Disorders
Mutations in genes encoding acyltransferases can cause metabolic disorders. For example, deficiency in carnitine acyltransferase leads to impaired fatty acid oxidation and cardiomyopathy. Likewise, acyl-CoA:cholesterol acyltransferase (ACAT) is involved in cholesterol esterification and atherosclerosis. Understanding the role of these enzymes in metabolism provides insights into disease mechanisms and potential therapies.
Acyltransferases in Infectious Diseases
Bacterial acyltransferases are essential for the biosynthesis of cell envelope components and virulence factors. For example, aminoglycoside acetyltransferases confer resistance to aminoglycoside antibiotics by acetylating the drug. Inhibitors of these enzymes could restore antibiotic sensitivity and are being explored as adjunct therapies. Additionally, mycobacterial acyltransferases involved in mycolic acid synthesis are targets for tuberculosis treatment.
From acyltransferase activity, transferring groups other than amino-acyl groups-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| What is the catalytic mechanism of ItaT? | Point mutation of active site residues in E. coli, kinetic assays |
| How does dehydrophos biosynthetic acyltransferase contribute to antibiotic production? | Knockout of biosynthetic genes in Streptomyces, metabolite profiling |
| Does histone acetyltransferase promote tumor growth? | Knockout in cancer cell lines, xenograft mouse models |
| Can acyltransferase inhibitors restore antibiotic sensitivity? | Overexpression of aminoglycoside acetyltransferase in resistant bacteria, MIC assays |
| What is the role of DGAT in lipid metabolism? | Knockout mice, overexpression in liver, lipidomics |
| How does carnitine acyltransferase deficiency affect heart function? | Knock-in mice with patient mutations, echocardiography |
How to Study the acyltransferase activity, transferring groups other than amino-acyl groups Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Enzymatic assay with radiolabeled donor | Acyltransferase activity | Kinetic characterization of purified enzymes |
| Mass spectrometry | Acylated product formation | Substrate specificity profiling |
| X-ray crystallography | Three-dimensional structure | Active site mapping and inhibitor design |
| CRISPR knockout screen | Gene essentiality and resistance | Identifying novel acyltransferases |
| RNA-seq | Transcriptional changes | Pathway analysis upon acyltransferase perturbation |
| Metabolomics | Metabolite levels | Detecting acylated metabolites |
| Western blot | Protein expression and modification | Validating acetylation events |
| Immunoprecipitation | Protein-protein interactions | Identifying acyltransferase complexes |
Biochemical Assays for Acyltransferase Activity
Direct measurement of acyltransferase activity typically involves incubating the enzyme with a donor and acceptor, followed by detection of the acylated product using chromatography or mass spectrometry. For example, ItaT activity can be assayed by monitoring the acetylation of aminoacyl-tRNAs using radiolabeled acetyl-CoA. These assays are essential for determining kinetic parameters and substrate specificity.
Structural Biology and Molecular Docking
X-ray crystallography and cryo-EM can provide atomic-level insights into the active site and substrate binding of acyltransferases. Molecular docking and molecular dynamics simulations complement experimental structures to predict how mutations affect catalysis. These methods guide the design of inhibitors and engineered enzymes.
CRISPR Screens to Identify Acyltransferase Functions
Genome-wide CRISPR knockout screens can systematically identify genes with acyltransferase activity that are required for specific cellular processes, such as drug resistance or metabolic adaptation. For instance, a screen for aminoglycoside resistance could reveal novel acetyltransferases. Similarly, CRISPR interference (CRISPRi) can be used to knockdown essential acyltransferases and study their roles.
Omics Approaches for Pathway Analysis
Transcriptomics, proteomics, and metabolomics can reveal the broader impact of acyltransferase activity on cellular metabolism. For example, RNA-seq of cells treated with acyltransferase inhibitors can identify downstream transcriptional changes. Metabolomics can detect altered levels of acylated metabolites, providing a functional readout.
How CRISPR Can Be Used to Study GO:0016747 acyltransferase activity, transferring groups other than amino-acyl groups
Knockout
CRISPR knockout of genes encoding acyltransferases can reveal their essentiality and cellular functions. For example, knocking out ItaT in E. coli abolishes aminoacyl-tRNA acetylation, leading to growth defects under specific conditions. Similarly, knockout of histone acetyltransferases in cancer cell lines can reduce proliferation and alter gene expression. These models are valuable for validating drug targets and understanding disease mechanisms.
Point Mutation
Introducing point mutations in the catalytic residues of acyltransferases via CRISPR can dissect their enzymatic mechanism. For instance, mutating the active site cysteine of ItaT abolishes its acetyltransferase activity, confirming its catalytic role. Such models are crucial for understanding structure-function relationships and for engineering enzymes with altered specificity.
Knock-in
Knock-in of disease-associated mutations in acyltransferase genes can create cellular and animal models of human disorders. For example, knocking in a patient mutation in carnitine acyltransferase into mice recapitulates cardiomyopathy phenotypes. These models are used to test therapeutic interventions and study disease progression.
Overexpression
Overexpression of acyltransferases can mimic gain-of-function states observed in cancer and metabolic diseases. For example, overexpressing DGAT in adipocytes increases triglyceride accumulation and lipid droplet formation. Overexpression models are also used to produce large quantities of enzymes for biochemical studies.
How EDITGENE Supports acyltransferase activity, transferring groups other than amino-acyl groups Research
Researchers studying acyltransferase activity, transferring groups other than amino-acyl groups-related genes often need to determine whether a candidate gene is causally involved in a specific pathway or disease. EDITGENE provides a comprehensive suite of CRISPR-based services to accelerate this discovery process, from gene knockout to precise point mutations and library screening.
Contact EDITGENE today to design your custom CRISPR model for acyltransferase activity, transferring groups other than amino-acyl groups research.
Frequently Asked Questions About acyltransferase activity, transferring groups other than amino-acyl groups
What is acyltransferase activity, transferring groups other than amino-acyl groups?
It is a molecular function defined by GO:0016747, describing enzymes that catalyze the transfer of an acyl group from a donor to an acceptor, excluding amino-acyl transfer.
What genes are involved in acyltransferase activity, transferring groups other than amino-acyl groups?
Genes include ItaT in E. coli, dehydrophos biosynthetic enzymes, histone acetyltransferases, diacylglycerol acyltransferases, and many others.
What diseases are associated with acyltransferase dysfunction?
Dysregulation is linked to cancer, metabolic disorders, and infectious diseases.
How can I study acyltransferase activity in the lab?
Common methods include enzymatic assays, mass spectrometry, structural biology, and CRISPR screens.
What is the difference between GO:0016747 and aminoacyltransferase activity?
GO:0016747 explicitly excludes the transfer of amino-acyl groups, which is covered by a different GO term.
Which model organisms are used to study acyltransferase activity?
E. coli, yeast, mice, and human cell lines are commonly used.
Can CRISPR be used to study acyltransferase function?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression are powerful tools for functional studies.
What are the substrates of acyltransferases?
Substrates vary widely and include acetyl-CoA, malonyl-CoA, aminoacyl-tRNAs, and lipids.
How is acyltransferase activity regulated?
Regulation occurs at transcriptional, post-translational, and allosteric levels.
What services does EDITGENE offer for acyltransferase research?
EDITGENE provides CRISPR knockout, point mutation, knock-in, overexpression, library screening, and bioinformatics services.
Conclusion
GO:0016747 acyltransferase activity, transferring groups other than amino-acyl groups, represents a diverse and biologically critical class of enzymes. From natural product biosynthesis to tRNA modification and disease pathology, these enzymes are central to many cellular processes. Advances in CRISPR-based models and biochemical assays continue to unravel their mechanisms and therapeutic potential. EDITGENE is committed to supporting this research with tailored CRISPR services, enabling precise genetic manipulation and functional discovery.
References
- 1. Ulrich EC et al.. 2018. Investigation of Amide Bond Formation during Dehydrophos Biosynthesis.. ACS Chem Biol 13(3):537-541 PMID: 29303545
- 2. Zhang C et al.. 2020. Substrate specificities of Escherichia coli ItaT that acetylates aminoacyl-tRNAs.. Nucleic Acids Res 48(13):7532-7544 PMID: 32501503
