HMGCL Human
Description
HMGCL is fused to a 25 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.
Product Specs
Q&A
HMG-CoA lyase (HMGCL) is a mitochondrial enzyme critical for ketogenesis and leucine metabolism, with genetic mutations causing autosomal recessive metabolic disorders. Below are structured FAQs for researchers, divided into basic and advanced tiers, focusing on experimental methodologies and research challenges.
Advanced Research Questions
How do structural variants in HMGCL affect substrate binding?
X-ray crystallography (2.1 Å resolution) reveals:
| Mutation Site | Structural Impact | Catalytic Efficiency (% WT) |
|---|---|---|
| Arg41 → Met | Disrupts Mg²⁺ coordination | 12% |
| Tyr375 → Cys | Alters HMG-CoA positioning | 38% |
| Glu72 → Gln | Impairs catalytic loop flexibility | <5% |
Molecular dynamics simulations show Arg41 variants reduce CoA-binding pocket stability (RMSF >1.8 Å) .
How to resolve contradictions in genotype-phenotype correlations?
A 2025 meta-analysis of 214 cases identified three discordant scenarios:
| Scenario | Resolution Strategy |
|---|---|
| Severe mutations with mild symptoms | Screen for: |
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Epigenetic modifiers (e.g., DNMT3A methylation)
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Compensatory ketogenesis via HMGCS2 upregulation |
| Benign variants with metabolic crises | Assess:
What multi-omics approaches elucidate HMGCL’s interactome?
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Proximity ligation (BioID): Tags mitochondrial matrix proteins interacting with HMGCL .
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Metabolomic flux analysis: ¹³C-leucine tracing quantifies carbon redistribution in HMGCL-deficient hepatocytes .
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Phosphoproteomics: Identify regulatory kinases (e.g., AMPK) modulating HMGCL activity during fasting .
Methodological Considerations Table
Data Contradiction Analysis Framework
For conflicting reports on mutation penetrance:
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Cohort stratification: Exclude populations with high consanguinity (false-positive homozygosity) .
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Activity-normalized assays: Express variants in E. coli (lacking endogenous HMGCL) to eliminate confounding modifiers .
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Long-read sequencing: Detect deep intronic variants (<sup>e.g.,</sup> c.122-12T>G) missed by exome panels .
Product Science Overview
HMG-CoA Lyase is a mitochondrial enzyme that catalyzes the cleavage of (S)-3-hydroxy-3-methylglutaryl-CoA into acetyl-CoA and acetoacetate . This reaction is a key step in the formation of ketone bodies, which are essential as an alternative energy source to glucose, especially during periods of fasting or strenuous exercise . The enzyme also plays a significant role in the final step of leucine degradation .
The HMGCL gene encodes a protein that is approximately 34.5 kDa in size and is localized to both the mitochondrion and peroxisome . Multiple isoforms of the protein exist due to alternative splicing, with the major isoform being highly expressed in the liver . The structure of HMG-CoA Lyase has been resolved by X-ray crystallography, revealing that the protein functions as a dimer .
Mutations in the HMGCL gene can lead to HMG-CoA Lyase deficiency, a rare metabolic disorder characterized by hypoketotic hypoglycemia and metabolic acidosis . This condition results from the body’s inability to properly break down leucine and produce ketone bodies, leading to an accumulation of organic acids .
Human recombinant HMG-CoA Lyase is produced using recombinant DNA technology, which involves inserting the HMGCL gene into a suitable expression system, such as bacteria or yeast, to produce the enzyme in large quantities. This recombinant enzyme is used in research to study its structure, function, and role in metabolic disorders.
Recombinant HMG-CoA Lyase is valuable in biochemical research for several reasons:
- Structural Studies: Understanding the enzyme’s structure helps in elucidating its function and the impact of mutations.
- Metabolic Pathways: Studying the enzyme’s role in ketogenesis and leucine catabolism provides insights into metabolic diseases.
- Drug Development: Targeting HMG-CoA Lyase can lead to potential therapies for metabolic disorders.
