GO:0102965 alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0102965 describes the NADPH-dependent reduction of a long-chain fatty acyl-CoA (13-22 carbons) to a long-chain primary fatty alcohol, releasing CoA and NADP+.
• This activity is a branch-point reaction that diverts fatty acyl-CoAs away from membrane phospholipid and triglyceride synthesis toward fatty alcohol production.
• The reaction requires two reducing equivalents from NADPH and proceeds via a fatty aldehyde intermediate, although the exact enzyme-bound mechanism remains under investigation.
• Enzyme activity can be measured in cell lysates or microsomal fractions using radiolabeled or fluorescent acyl-CoA substrates and NADPH.
• Defects in fatty alcohol metabolism are linked to peroxisomal disorders such as Zellweger syndrome, where acyl-CoA reductase activities are altered.
• CRISPR knockout, point-mutation, and overexpression models enable causal testing of candidate genes annotated to GO:0102965 in human cells.
Description
GO:0102965, alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity, is a molecular function that catalyzes the NADPH-dependent conversion of a long-chain fatty acyl-CoA into a long-chain primary fatty alcohol, CoA, and NADP+. Long-chain fatty acids in this context contain aliphatic tails of 13 to 22 carbons, placing this activity at the intersection of fatty acid metabolism and the biosynthesis of fatty alcohols, which serve as precursors for wax esters and ether lipids. Researchers study this activity because it represents a key branch point that determines whether acyl-CoAs are channeled into energy storage, membrane biogenesis, or specialized lipid products. In cultured fibroblasts from Zellweger syndrome patients, enzymes catalyzing the biosynthesis of lysophosphatidate and its ether analog show altered properties, highlighting the importance of peroxisomal and microsomal fatty acyl-CoA reductases in human lipid homeostasis. Understanding GO:0102965 therefore has implications for peroxisomal disorders, lipid metabolic engineering, and the development of cell models to dissect gene function.
alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity At A Glance
| GO ID | GO:0102965 |
|---|---|
| GO term | alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity |
| Ontology | molecular_function |
| Synonym | alcohol-forming fatty acyl-CoA reductase activity |
| Definition | Catalysis of the reaction: a long-chain fatty acyl-CoA + 2 H+ + 2 NADPH = a long-chain primary fatty alcohol + CoA + 2 NADP+. A long-chain fatty acid has an aliphatic tail containing 13 to 22 carbons. |
| Substrate | Long-chain fatty acyl-CoA (13-22 carbons) |
| Cofactor | NADPH (two molecules per reaction) |
| Products | Long-chain primary fatty alcohol, CoA, NADP+ |
| Reaction direction | Reductive; consumes NADPH and acyl-CoA |
What Is GO:0102965?
GO:0102965 is defined as the catalysis of the reaction: a long-chain fatty acyl-CoA + 2 H+ + 2 NADPH = a long-chain primary fatty alcohol + CoA + 2 NADP+. A long-chain fatty acid has an aliphatic tail containing 13 to 22 carbons. In other words, this activity reduces the thioester carbonyl of a fatty acyl-CoA to a primary alcohol using NADPH as the electron donor, releasing coenzyme A and oxidized NADP+.
Why Is alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity Important in Cell Biology?
GO:0102965 is important because it controls the flux of long-chain fatty acyl-CoAs into fatty alcohol production, a pathway that supplies precursors for wax esters, ether lipids, and signaling molecules. In humans, perturbations in fatty alcohol metabolism are associated with peroxisomal disorders such as Zellweger syndrome, where enzyme activities involved in lysophosphatidate and ether lipid biosynthesis are abnormal. Studying this activity helps researchers understand how cells balance membrane lipid synthesis, energy storage, and specialized lipid mediator production, and it provides a target for metabolic engineering and therapeutic intervention.
• Controls a branch point between membrane phospholipid synthesis and fatty alcohol production.
• Provides precursors for wax esters and ether lipids, which are important for membrane structure and signaling.
• Altered activity is observed in peroxisomal disorders such as Zellweger syndrome.
• Relevant to metabolic engineering of fatty alcohol production in microbial and mammalian cells.
• Serves as a model for studying NADPH-dependent redox reactions in lipid metabolism.
• Can be targeted by CRISPR to test causal roles in lipid homeostasis.
• Links fatty acid metabolism to cellular detoxification and lipid signaling.
• Potential biomarker or therapeutic target in disorders of lipid metabolism.
What Happens During alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity?
Substrate binding and activation
In simple terms: The enzyme grabs a fatty acyl-CoA molecule and prepares it for chemical modification.
The reaction begins with the binding of a long-chain fatty acyl-CoA (13-22 carbons) to the active site of the enzyme. The thioester bond between the fatty acid and coenzyme A is polarized, making the carbonyl carbon susceptible to reduction. This step is essential for positioning the substrate for the subsequent transfer of hydride equivalents from NADPH.
NADPH-dependent reduction to fatty aldehyde
In simple terms: NADPH donates electrons to convert the fatty acyl-CoA into a fatty aldehyde intermediate.
The enzyme catalyzes the transfer of hydride from NADPH to the thioester carbonyl, forming a fatty aldehyde intermediate. This reductive step releases coenzyme A and consumes one molecule of NADPH, generating NADP+. The aldehyde intermediate remains bound to the enzyme or is released into the membrane for further reduction.
Second reduction to primary fatty alcohol
In simple terms: A second round of reduction converts the aldehyde into a fatty alcohol.
A second molecule of NADPH reduces the fatty aldehyde to a long-chain primary fatty alcohol. This step completes the four-electron reduction of the acyl-CoA to an alcohol, releasing NADP+ and the final product. The overall reaction stoichiometry is: long-chain fatty acyl-CoA + 2 H+ + 2 NADPH = long-chain primary fatty alcohol + CoA + 2 NADP+.
Product release and metabolic fate
In simple terms: The fatty alcohol is released and can be used to build waxes, ether lipids, or other molecules.
The newly synthesized fatty alcohol is released from the enzyme and can be further metabolized into wax esters, ether lipids, or other specialized lipids. In mammalian cells, these products contribute to membrane structure and signaling. The release of CoA and NADP+ regenerates cofactors for other metabolic reactions.
Key Genes Involved in GO:0102965 alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity
The following genes and proteins are associated with or relevant to GO:0102965, based on published literature and functional annotations.
| Gene | Major Role | Research Relevance |
|---|---|---|
| FAR1 | Fatty acyl-CoA reductase 1 | Catalyzes alcohol-forming long-chain fatty acyl-CoA reductase activity; implicated in wax ester synthesis. |
| FAR2 | Fatty acyl-CoA reductase 2 | Contributes to fatty alcohol production in peroxisomes; linked to ether lipid synthesis. |
| ACSL1 | Acyl-CoA synthetase long-chain family member 1 | Generates long-chain fatty acyl-CoA substrates for GO:0102965. |
| ACSL3 | Acyl-CoA synthetase long-chain family member 3 | Provides acyl-CoA substrates in lipid droplets and membranes. |
| ACSL4 | Acyl-CoA synthetase long-chain family member 4 | Supplies acyl-CoAs for reductive pathways including fatty alcohol synthesis. |
| GPAT1 | Glycerol-3-phosphate acyltransferase 1 | Competes for acyl-CoA substrates, influencing flux through GO:0102965. |
| AGPAT2 | 1-acylglycerol-3-phosphate O-acyltransferase 2 | Competes for acyl-CoA in phospholipid synthesis, affecting fatty alcohol production. |
| DGAT1 | Diacylglycerol O-acyltransferase 1 | Diverts acyl-CoA to triglycerides, indirectly regulating GO:0102965 flux. |
| DGAT2 | Diacylglycerol O-acyltransferase 2 | Similar to DGAT1, affects acyl-CoA partitioning. |
| PEX5 | Peroxisomal biogenesis factor 5 | Mutations cause Zellweger syndrome; peroxisomal fatty alcohol metabolism is altered. |
| PEX1 | Peroxisomal biogenesis factor 1 | Defects lead to peroxisomal disorders with abnormal lipid metabolism. |
| PEX6 | Peroxisomal biogenesis factor 6 | Involved in peroxisome assembly; relevant to fatty alcohol metabolism. |
| ABCD1 | ATP binding cassette subfamily D member 1 | X-linked adrenoleukodystrophy gene; affects very long-chain fatty acid metabolism. |
| HSD17B4 | Hydroxysteroid 17-beta dehydrogenase 4 | Peroxisomal multifunctional enzyme involved in fatty acid oxidation. |
| ACOX1 | Acyl-CoA oxidase 1 | Peroxisomal fatty acid oxidation; influences acyl-CoA pools. |
| CYP4A11 | Cytochrome P450 family 4 subfamily A member 11 | Omega-hydroxylase that can produce fatty alcohols from fatty acids. |
| FAR2P1 | Fatty acyl-CoA reductase 2 pseudogene 1 | Potential regulatory role; requires experimental validation. |
How Is alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity Regulated?
The activity of alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) is regulated at multiple levels. Substrate availability of long-chain fatty acyl-CoAs, generated by acyl-CoA synthetases, directly influences flux through the reaction. NADPH supply from the pentose phosphate pathway and malic enzyme provides reducing equivalents required for catalysis. In peroxisomal disorders such as Zellweger syndrome, defects in peroxisome biogenesis alter the enzymatic environment and can affect the properties of enzymes catalyzing lysophosphatidate and ether lipid biosynthesis, which are metabolically linked to fatty alcohol production. Transcriptional regulation of FAR1 and FAR2 genes by lipid-sensing nuclear receptors further modulates activity.
alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| PEX5 | Zellweger syndrome | Patient-derived fibroblasts with PEX5 mutations; CRISPR KO in HEK293 |
| ABCD1 | X-linked adrenoleukodystrophy | ABCD1 knockout iPSC-derived oligodendrocytes |
| FAR1 | Fatty alcohol metabolic disorders | FAR1 knockout HeLa cells; overexpression in HEK293 |
| FAR2 | Peroxisomal ether lipid deficiency | FAR2 knockout HepG2 cells |
| ACSL4 | Ferroptosis and lipid peroxidation | ACSL4 knockout cancer cell lines |
Zellweger syndrome and peroxisomal disorders
Zellweger syndrome is a peroxisomal biogenesis disorder characterized by severe neurological impairment and lipid metabolic abnormalities. Cultured fibroblasts from Zellweger syndrome patients show altered properties of enzymes catalyzing the biosynthesis of lysophosphatidate and its ether analog, indicating that peroxisomal dysfunction impacts lipid pathways connected to fatty alcohol metabolism. GO:0102965 activity may be indirectly affected due to disrupted peroxisomal fatty acid handling.
X-linked adrenoleukodystrophy
X-linked adrenoleukodystrophy is caused by mutations in ABCD1, leading to accumulation of very long-chain fatty acids. This accumulation can alter acyl-CoA pools and potentially affect the flux through reductive pathways such as GO:0102965, contributing to the pathology of the disease.
Metabolic syndrome and lipid storage disorders
Dysregulation of fatty acyl-CoA partitioning between oxidation, storage, and fatty alcohol synthesis may contribute to metabolic disorders. Enzymes competing for acyl-CoA substrates, such as GPAT1 and DGATs, influence the availability of substrates for GO:0102965, linking this activity to lipid storage diseases.
From alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of FAR1 reduce long-chain fatty alcohol production? | FAR1 knockout HEK293 cells generated by CRISPR |
| Does a point mutation in the NADPH-binding domain abolish activity? | Point-mutation knock-in of FAR1 catalytic residues |
| Can tagged FAR1 be used to monitor subcellular localization? | Knock-in of FLAG- or GFP-tagged FAR1 |
| Does overexpression of FAR2 increase ether lipid synthesis? | FAR2 overexpression in HeLa cells |
| Which genes regulate flux through GO:0102965? | CRISPR library screening with fatty alcohol reporters |
| Is GO:0102965 activity altered in Zellweger syndrome? | Patient fibroblasts and PEX5 knockout models |
How to Study the alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Radiolabeled acyl-CoA assay | Enzymatic conversion to fatty alcohol | Validation of knockout or point-mutation effects |
| LC-MS lipidomics | Fatty alcohol and wax ester levels | Profiling metabolic flux in cells |
| RNA-seq | Transcript levels of FAR and related genes | Identifying regulatory changes |
| Proteomics | Protein abundance and modifications | Detecting post-translational regulation |
| Fluorescence microscopy | Subcellular localization of tagged enzymes | Determining organelle-specific activity |
| CRISPR library screening | Genes affecting fatty alcohol production | Discovery of novel regulators |
| Co-immunoprecipitation | Protein-protein interactions | Identifying enzyme complexes |
| NADPH consumption assay | Redox cofactor usage | Kinetic characterization |
Enzymatic activity assays
Alcohol-forming long-chain fatty acyl-CoA reductase activity can be measured in cell lysates or microsomal fractions using radiolabeled or fluorescent long-chain acyl-CoA substrates and NADPH. The formation of fatty alcohol products is quantified by thin-layer chromatography, HPLC, or mass spectrometry. These assays are essential for validating CRISPR knockout or point-mutation effects on GO:0102965.
Lipidomics and metabolomics
Mass spectrometry-based lipidomics enables comprehensive profiling of fatty alcohols, wax esters, and ether lipids in cells and tissues. This approach can detect changes in metabolic flux through GO:0102965 in response to genetic perturbations or drug treatments.
Transcriptomics and proteomics
RNA-seq and quantitative proteomics can reveal changes in the expression of genes encoding fatty acyl-CoA reductases and related enzymes. Integrating these datasets with CRISPR screens helps identify regulatory networks controlling GO:0102965 activity.
Imaging and subcellular localization
Fluorescence microscopy of tagged FAR proteins or lipid dyes can visualize the subcellular compartments where fatty alcohol synthesis occurs, such as the endoplasmic reticulum and peroxisomes. This is particularly relevant for understanding how peroxisomal disorders affect GO:0102965.
How CRISPR Can Be Used to Study GO:0102965 alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity
Knockout
CRISPR knockout of candidate genes such as FAR1 or FAR2 can abolish or reduce alcohol-forming long-chain fatty acyl-CoA reductase activity, providing causal evidence for their role in GO:0102965. Knockout cell lines are generated by introducing indels in early exons, followed by functional validation using enzymatic assays or lipidomics.
Point Mutation
Point mutations in catalytic residues or NADPH-binding motifs can be introduced via CRISPR base editing or homology-directed repair to dissect the mechanism of GO:0102965. Such models help distinguish between loss-of-function, hypomorphic, and separation-of-function alleles.
Knock-in
Knock-in of epitope tags (e.g., FLAG, GFP) or reporter cassettes allows real-time monitoring of enzyme localization and expression. Tagged knock-in models are valuable for studying the subcellular site of fatty alcohol synthesis and for affinity purification of the enzyme complex.
Overexpression
CRISPR activation or lentiviral overexpression of FAR genes can increase GO:0102965 activity, enabling gain-of-function studies. Overexpression models are useful for testing whether increased fatty alcohol production affects membrane composition, signaling, or disease phenotypes.
How EDITGENE Supports alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity Research
Researchers studying alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity-related genes often need to determine whether a candidate gene is causally involved in fatty alcohol synthesis or merely correlated with changes in lipid metabolism. EDITGENE provides a comprehensive suite of CRISPR-based services to generate precisely engineered cell models, enabling rigorous functional validation of genes annotated to GO:0102965.
Contact EDITGENE today to design your custom CRISPR model for alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity research.
Frequently Asked Questions About alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity
What is GO:0102965?
GO:0102965 is a Gene Ontology molecular function term describing alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity, which catalyzes the NADPH-dependent reduction of a long-chain fatty acyl-CoA to a long-chain primary fatty alcohol, CoA, and NADP+.
What reaction does alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity catalyze?
It catalyzes: a long-chain fatty acyl-CoA + 2 H+ + 2 NADPH = a long-chain primary fatty alcohol + CoA + 2 NADP+, where the fatty acid has 13 to 22 carbons.
What genes are involved in alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity?
Genes such as FAR1, FAR2, and acyl-CoA synthetases (ACSL1, ACSL3, ACSL4) are involved in supplying substrates or catalyzing the reaction.
What is the synonym for GO:0102965?
The synonym is alcohol-forming fatty acyl-CoA reductase activity.
How is alcohol-forming long-chain fatty acyl-CoA reductase activity measured?
It is measured using enzymatic assays with radiolabeled or fluorescent acyl-CoA substrates and NADPH, followed by product detection via chromatography or mass spectrometry.
Is GO:0102965 associated with any diseases?
Altered fatty alcohol metabolism has been observed in peroxisomal disorders such as Zellweger syndrome, where enzymes involved in related lipid pathways show abnormal properties.
What cofactor does GO:0102965 require?
It requires NADPH as the electron donor, consuming two molecules per reaction.
What is the subcellular localization of alcohol-forming long-chain fatty acyl-CoA reductase?
The activity is associated with the endoplasmic reticulum and peroxisomes, depending on the specific enzyme isoform.
Can CRISPR be used to study GO:0102965?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models enable causal testing of genes involved in this activity.
What are the products of the GO:0102965 reaction?
The products are a long-chain primary fatty alcohol, coenzyme A, and NADP+.
Conclusion
GO:0102965, alcohol-forming long-chain fatty acyl-CoA reductase (NADP+) activity, is a key molecular function that links fatty acyl-CoA metabolism to the production of long-chain fatty alcohols, precursors for wax esters and ether lipids. Its relevance to peroxisomal disorders such as Zellweger syndrome underscores its importance in human health. By combining precise CRISPR engineering with biochemical and lipidomic assays, researchers can dissect the genetic and mechanistic basis of this activity, paving the way for therapeutic and biotechnological applications.
References
- 1. Webber KO et al.. 1987. Properties of the enzymes catalyzing the biosynthesis of lysophosphatidate and its ether analog in cultured fibroblasts from Zellweger syndrome patients and normal controls.. Arch Biochem Biophys 254(2):611-20 PMID: 3646870
