LANCL1 Human
Description
Biological Roles and Mechanisms
LANCL1 regulates redox homeostasis and cellular signaling through distinct mechanisms:
Redox Regulation and CBS Interaction
LANCL1 binds cystathionine β-synthase (CBS), inhibiting its activity under basal conditions. Oxidative stress disrupts this interaction, releasing CBS to enhance cysteine/GSH synthesis :
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Mechanism: GSH/GSSG ratio modulates LANCL1-CBS binding.
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Outcome: Elevated CBS activity compensates for reduced GSH during oxidative stress .
Neuroprotection in ALS
In amyotrophic lateral sclerosis (ALS) models:
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Transgenic Overexpression: Prolongs lifespan, delays disease onset, and improves motor function in SOD1 G93A mice .
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AKT Pathway Activation: Restores phosphorylation of AKT, mitigating oxidative damage and neuroinflammation .
Cancer Progression
LANCL1 promotes prostate cancer cell proliferation and protects against oxidative stress:
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JNK Pathway Inhibition: Suppresses apoptosis and enhances survival .
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Clinical Correlation: Upregulated in advanced prostate tumors .
Protein Interactions and Signaling
LANCL1 interacts with diverse partners, influencing cytoskeletal dynamics and signaling:
Key Interaction Partners
Neurodegenerative Diseases
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ALS: CNS-specific deletion of LANCL1 exacerbates motor neuron loss and oxidative damage, while transgenic overexpression alleviates symptoms .
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Oxidative Stress: Acts as a redox sensor, modulating CBS activity to maintain GSH levels .
Cancer Therapy
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Target Potential: Knockdown of LANCL1 reduces prostate cancer cell survival and apoptosis resistance .
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Biomarker Utility: Elevated LANCL1 correlates with tumor progression in prostate cancer .
Future Research Directions
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Mechanistic Studies: Elucidate the role of LANCL1 in AKT activation and cytoskeletal dynamics.
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Therapeutic Strategies: Explore LANCL1 targeting in ALS and cancer, leveraging its redox-modulating properties.
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Structural Dynamics: Investigate conformational changes during GSH binding and CBS interaction.
Product Specs
Q&A
What is LANCL1 and what is its primary function in human physiology?
LANCL1 is a member of the lanthionine synthetase C-like protein family that functions as a receptor for abscisic acid (ABA), a molecule originally identified as a plant hormone but now recognized as active in animals as well. LANCL1 shares significant sequence identity (54.2%) with its paralog LANCL2, and both are attached to the inner side of the plasma membrane .
Unlike their bacterial homologs that synthesize lanthionine-containing peptides with antibiotic properties, mammalian LANCL proteins have evolved different functions. LANCL1 primarily regulates glucose metabolism by stimulating insulin-independent glucose uptake through activation of glucose transporters. It also enhances mitochondrial function and energy metabolism via the AMPK/PGC-1α/Sirt1 signaling pathway . The physiological importance of LANCL proteins is underscored by the finding that triple LANCL knockout mice die prematurely .
How does LANCL1 interact with abscisic acid and what binding affinity has been measured?
LANCL1 binds ABA with a dissociation constant (Kd) between 1 and 10 μM, depending on the assay methodology . This binding affinity lies between the low and high-affinity ABA binding sites of LANCL2. The interaction has been characterized using multiple complementary techniques:
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Equilibrium-binding experiments with [³H]ABA
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Circular dichroism spectroscopy
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Surface plasmon resonance
These methods have collectively established LANCL1 as a genuine ABA receptor, though with somewhat lower affinity than LANCL2 . For researchers investigating this interaction, it's important to use multiple binding assay approaches to fully characterize the binding parameters, as different methodologies may yield slightly different Kd values within the micromolar range.
What is the relationship between LANCL1 and LANCL2 expression in tissues?
LANCL1 and LANCL2 exhibit a fascinating compensatory relationship in mammalian tissues. When one receptor is downregulated or absent, the other is spontaneously upregulated:
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LANCL2−/− mice spontaneously overexpress LANCL1 in skeletal muscle
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Silencing of LANCL2 in cells induces overexpression of LANCL1
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Conversely, silencing of LANCL1 results in overexpression of LANCL2 in adipose tissue and muscle cells
This reciprocal regulation suggests an important physiological redundancy mechanism that maintains ABA signaling. Both proteins are similarly and ubiquitously expressed across mammalian tissues and organs in normal conditions . This redundancy appears evolutionarily conserved, likely due to the critical role of the ABA/LANCL system in controlling insulin-independent glucose uptake and energy metabolism .
What experimental approaches are most effective for studying LANCL1-mediated glucose transport?
To investigate LANCL1's effect on glucose transport, researchers have successfully employed rat L6 myoblast models with genetic manipulation of LANCL1 expression. The following methodological approach has proven effective:
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Generate cell lines overexpressing LANCL1 or with LANCL1/LANCL2 silencing via appropriate vectors
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Validate protein expression levels using Western blot and qPCR (typical overexpression levels are 10-40 times higher than control cells)
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Measure glucose uptake using fluorescent glucose analogs like NBDG
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Analyze expression of glucose transporters (GLUT1, GLUT4) via qPCR and Western blot
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Assess downstream signaling through the AMPK/PGC-1α/Sirt1 pathway
In LANCL1-overexpressing myoblasts, researchers can expect to observe approximately 4-fold increases in both basal and ABA-stimulated glucose uptake compared to control cells . Similarly, 4-6 fold increases in GLUT4 and GLUT1 expression should be detectable, providing clear measurable outcomes for experimental manipulation.
How can researchers quantify LANCL1's role in mitochondrial function and thermogenesis?
LANCL1's impact on mitochondrial function and heat production can be quantified using several complementary approaches:
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Oxygen consumption measurements: Assess mitochondrial respiration rates in cells with modified LANCL1 expression. LANCL1 overexpression typically increases oxygen consumption approximately 5-fold .
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Direct heat production measurement: Two effective methods have been validated:
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Differential temperature measurement using super-sensitive temperature probes
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Differential scanning calorimetry (DSC)
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Uncoupling protein expression analysis: Measure expression of uncoupling proteins like UCP3 (increased ~12-fold with LANCL1 overexpression) and sarcolipin (increased ~3-fold) .
When measuring heat production in LANCL1/2-overexpressing versus double-silenced cells, researchers can expect to observe approximately double the heat production in overexpressing cells . Treatment with electron transfer chain inhibitors (rotenone and antimycin A) nearly abolishes this difference, confirming the mitochondrial origin of the heat generation .
What are the methodological considerations for studying ABA-LANCL1 signaling in vivo?
When investigating ABA-LANCL1 signaling in vivo, several methodological considerations are critical:
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Genetic models: LANCL2−/− mice provide a valuable model to study LANCL1 function in isolation, as they spontaneously overexpress LANCL1 in skeletal muscle .
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ABA administration protocol: Chronic ABA treatment at 1 μg/kg body weight/day has been established as effective .
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Functional readouts:
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Glucose tolerance tests to assess glycemic response
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Tissue collection for molecular analysis of the AMPK/PGC-1α/Sirt1 pathway
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Analysis of glucose transporter expression in skeletal muscle
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Expected outcomes: In LANCL2−/− mice treated with ABA, researchers should expect:
What approaches can be used to differentiate between LANCL1 and LANCL2 functions in experimental systems?
Differentiating between the functions of these similar receptors requires careful experimental design:
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Gene-specific silencing: Use siRNA or shRNA targeting specific regions of LANCL1 or LANCL2 that do not share sequence homology.
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Selective overexpression: Create cell lines overexpressing one receptor at a time to study its specific effects.
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Knockout models: Compare phenotypes of LANCL1−/−, LANCL2−/−, and wild-type animals.
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Comparative binding studies: Assess ABA binding affinities to distinguish receptor-specific dynamics:
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Rescue experiments: In LANCL2−/− models, introduce exogenous LANCL1 to assess functional rescue.
While both receptors activate similar pathways, their different binding affinities for ABA may result in distinct activation thresholds and potentially different physiological responses at varying ABA concentrations.
What are the current technical challenges in measuring cellular heat production mediated by LANCL1/2?
Measuring cellular thermogenesis presents several technical challenges:
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Small sample volumes: Working with approximately 2.5 × 10^6 cells in volumes of 100-150 μL requires highly sensitive instrumentation .
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Baseline temperature fluctuations: Initial temperature differences between measurement vials can range from 0.05-0.475°C, necessitating appropriate controls and equilibration periods .
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Signal interpretation: Distinguishing between thermal relaxation and cellular heat production requires mathematical modeling. The typical timescale for cells to metabolically "turn on" after addition of nutrients is approximately 4-7 minutes, which must be accounted for in experimental design .
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Calibration challenges: Experimental calibration is more reliable than theoretical models for estimating heating power, with typical values of 8-15 μW for LANCL1/2-overexpressing cells .
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Distinguishing sources of heat: To confirm mitochondrial origin of heat production, electron transport chain inhibitors should be included as controls.
Despite these challenges, differential temperature measurement and differential scanning calorimetry have successfully demonstrated that LANCL1/2-overexpressing cells produce approximately twice the heat of double-silenced cells, with a further 100% increase upon ABA treatment .
What are promising areas for expanding our understanding of LANCL1 function?
Several research directions hold potential for advancing LANCL1 research:
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Tissue-specific functions: While current research has focused on myocytes and adipocytes, exploring LANCL1 function in other tissues could reveal new physiological roles.
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Signaling pathway integration: Further investigation of how the AMPK/PGC-1α/Sirt1 pathway activated by LANCL1 integrates with other metabolic signaling networks.
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Evolutionary significance: Exploring why receptor redundancy between LANCL1 and LANCL2 has been maintained throughout evolution could reveal additional functions.
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ABA-independent functions: Investigating whether LANCL1 has physiological roles independent of ABA binding.
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Structural biology approaches: Determining the crystal structure of LANCL1 bound to ABA would provide valuable insights into binding mechanisms and potential for targeted modulation.
The redundancy and compensatory relationship between LANCL1 and LANCL2 suggest critical physiological importance of this signaling system, warranting continued investigation into its fundamental biology and potential therapeutic applications.
Product Science Overview
The LANCL1 gene encodes a peripheral membrane protein that is loosely associated with the cell membrane. This protein is related to the LanC family of bacterial membrane-associated proteins, which play a crucial role in the biosynthesis of lantibiotics, a class of antimicrobial peptides . The human recombinant form of LANCL1 is produced in Escherichia coli and consists of a single, non-glycosylated polypeptide chain containing 422 amino acids, with a molecular mass of approximately 47.7 kDa .
LANCL1 functions as a peptide-modifying enzyme component in eukaryotic cells. It is involved in the conjugation of glutathione (GSH) to artificial substrates such as 1-chloro-2,4-dinitrobenzene (CDNB) and p-nitrophenyl acetate. This activity suggests that LANCL1 plays a role in the glutathione antioxidant defense mechanism, helping to mitigate neuronal oxidative stress during normal postnatal development and in response to oxidative stresses .
LANCL1 is a 399-residue protein that was originally discovered in a search for binding partners to the erythrocyte protein stomatin, which is a factor in the disease hereditary overhydrated stomatocytosis . The protein binds glutathione and participates in EPS8 signaling, which is crucial for various cellular processes .
Mutations or dysregulation of the LANCL1 gene have been associated with several disorders, including intellectual developmental disorder, autosomal recessive 18, with or without epilepsy, and speech and communication disorders . The protein’s role in mitigating oxidative stress also suggests potential therapeutic applications in neurodegenerative diseases and other conditions characterized by oxidative damage.
The human recombinant form of LANCL1 is produced in Escherichia coli and is available as a sterile, filtered colorless solution. It is formulated in a buffer containing 20 mM Tris-HCl (pH 7.5), 0.15 M NaCl, 20% glycerol, and 1 mM DTT. For long-term storage, it is recommended to store the protein at -20°C with a carrier protein to avoid multiple freeze-thaw cycles .
