Recombinant Candida glabrata 3-keto-steroid reductase (ERG27)

What is the primary function of 3-keto-steroid reductase (ERG27) in C. glabrata?

ERG27 is a crucial enzyme in the ergosterol biosynthesis pathway of C. glabrata. It functions primarily as a steroid-3-ketoreductase, catalyzing the reduction of 3-ketosteroid intermediates in the pathway leading to ergosterol production. Ergosterol is the fungal equivalent of cholesterol and is essential for membrane integrity and cellular function. ERG27 has also been identified as a "moonlighting protein" with secondary functions beyond its catalytic role .

The enzyme is particularly notable for its involvement in the functionality of oxidosqualene cyclase (OSC, also known as Erg7p), which produces the first cyclical steroid intermediate in the pathway. Yeast cells lacking the ERG27 gene show concomitant loss of OSC activity and complete ablation of sterol synthesis, demonstrating the critical importance of this enzyme in fungal viability .

How does ERG27 differ structurally and functionally between C. glabrata and related Candida species?

C. glabrata ERG27 shares functional similarity with ERG27 homologs in related species like C. albicans, but with distinct evolutionary adaptations. While both participate in ergosterol biosynthesis, C. glabrata has evolved specific regulatory mechanisms that allow it to adapt to different host and environmental conditions .

Research comparing ERG27 across Candida species has revealed that while the catalytic domains are conserved, C. glabrata ERG27 has evolved specific protein-protein interaction interfaces that contribute to its dual functionality in both steroid reduction and OSC stabilization. Unlike some other Candida species, C. glabrata's ERG27 appears to be more tightly integrated with stress response pathways, potentially contributing to its enhanced antifungal resistance profile .

What are the consequences of ERG27 deletion in C. glabrata?

Deletion of ERG27 in C. glabrata leads to profound biological effects:

• Complete loss of ergosterol biosynthesis, as the pathway is interrupted

• Concomitant loss of oxidosqualene cyclase (OSC/Erg7p) activity

• Accumulation of pathway intermediates, particularly 3-ketosteroids

• Altered membrane composition and fluidity

• Growth arrest under sterol-deficient conditions

These effects demonstrate that ERG27 is essential for both its direct catalytic function and its role in supporting OSC activity. Experiments have shown that yeast mutants lacking ERG27 cannot grow on sterol-deficient media, highlighting the enzyme's essential nature . Interestingly, a catalytically inactive ERG27 can still retain its OSC protective function, resulting in the accumulation of 3-ketosteroids but allowing some cellular viability, which suggests distinct functional domains within the protein .

What are the optimal methods for heterologous expression of recombinant C. glabrata ERG27?

Based on successful expression systems reported in the literature, the optimal approach for recombinant C. glabrata ERG27 expression involves:

• Expression System Selection: E. coli has been successfully used for expressing the soluble fraction of related enzymes from C. glabrata, such as HMGR . For full-length ERG27 with membrane domains, yeast expression systems (particularly S. cerevisiae) often provide better results.

• Fusion Tag Strategy: Fusion to maltose binding protein (MBP) has proven effective for enhancing solubility and facilitating purification, as demonstrated with similar C. glabrata enzymes . Other commonly used tags include His6, GST, and SUMO.

• Expression Conditions:

  • Induction at OD600 of 0.6-0.8

  • IPTG concentration of 0.1-0.5 mM

  • Post-induction growth at 25-30°C rather than 37°C to enhance proper folding

  • Extended expression time (16-20 hours)

• Purification Protocol:

  • Initial capture using affinity chromatography based on the fusion tag

  • Ion exchange chromatography for further purification

  • Size exclusion chromatography as a final polishing step

For functional studies, it's critical to consider that ERG27 interacts with multiple proteins in the ergosterol pathway, so co-expression with binding partners may be necessary to maintain native conformation and activity .

How can researchers accurately measure ERG27 enzymatic activity in vitro?

Accurate measurement of C. glabrata ERG27 enzymatic activity requires:

• Substrate Preparation: Appropriate 3-ketosteroid substrates must be prepared or obtained. These can include physiologically relevant intermediates like 4-methylzymosterone, 4-methylfecosterone, and 4-methylepisterone (collectively called "3-ketosteroid 1" in some research) .

• Activity Assay Conditions:

  • Optimal pH of approximately 8.0 (based on similar C. glabrata enzymes)

  • Temperature of 37°C for maximum activity

  • NADPH as the cofactor (typically 100-200 μM)

  • Buffer systems containing phosphate or Tris-HCl

• Detection Methods:

  • Spectrophotometric monitoring of NADPH oxidation at 340 nm

  • TLC separation of substrate and product, especially useful when working with radiolabeled substrates

  • GC-MS confirmation of reaction products

• Validation Approaches:

  • NaBH4 treatment as a control to confirm 3-keto group reduction

  • Comparison with known inhibitors or with enzyme prepared from wild-type strains

For kinetic analysis, researchers should determine Km and Vmax values under varying substrate concentrations while maintaining excess cofactor. Based on data from related enzymes in C. glabrata, expected Km values may be in the low micromolar range (e.g., 6.5 μM for HMGR) .

What genetic tools and approaches are most effective for studying ERG27 function in C. glabrata?

Effective genetic approaches for studying ERG27 function include:

• Gene Deletion Strategies:

  • CRISPR-Cas9 systems adapted for C. glabrata

  • Homologous recombination using NAT1, KanMX, or HygB resistance markers

  • Conditional expression systems (tetracycline-regulated or GAL-inducible)

• Complementation Assays:

  • Expression of wild-type or mutant ERG27 in deletion strains

  • Cross-species complementation (testing mammalian homologs in ERG27-deficient yeast)

  • Domain swapping to identify functional regions

• Reporter Systems:

  • Fusion of ERG27 to GFP or other fluorescent proteins for localization studies

  • Luciferase reporters for studying transcriptional regulation

  • Split-protein assays for detecting protein-protein interactions

• Phenotypic Analyses:

  • Growth assays on different media (with/without sterol supplementation)

  • Antifungal susceptibility testing

  • Sterol profiling by GC-MS or HPLC

The ERG27 deletion can be complemented with various constructs to study specific functions. For example, researchers have demonstrated that mammalian HSD17B7 genes express poorly in yeast ERG27-deficient backgrounds, providing insights into structural and functional requirements for ERG27 activity .

How does ERG27 contribute to azole resistance mechanisms in C. glabrata?

ERG27 contributes to azole resistance in C. glabrata through several mechanisms:

Unlike some resistance mechanisms that directly involve drug transporters (such as CDR1, PDH1, and SNQ2), ERG27's contribution to resistance primarily involves maintaining ergosterol biosynthesis pathway integrity when other components are inhibited by antifungals .

Can ERG27 mutations confer resistance to polyene antifungals like amphotericin B?

While direct ERG27 mutations conferring polyene resistance haven't been extensively characterized, the enzyme plays an important indirect role in polyene susceptibility:

• Sterol Composition Effects: ERG27 disruption alters the sterol composition of the fungal membrane. Polyenes like amphotericin B target ergosterol directly, so changes in sterol content can affect drug binding and efficacy .

• Pathway Interactions: ERG27 interacts functionally with other ERG pathway enzymes. Research has shown that mutations in other ERG genes (like ERG2) can lead to reduced sensitivity to amphotericin B by changing membrane sterol composition .

• Compensatory Mechanisms: When ERG27 function is compromised, accumulation of alternative sterols may occur, potentially reducing polyene binding sites in the membrane.

In C. glabrata clinical isolates with reduced amphotericin B sensitivity (MIC of 8 μg/ml), mutations have been identified in ERG2, leading to accumulation of Δ8-sterol intermediates . While these studies don't directly implicate ERG27 mutations, they demonstrate how alterations in the ergosterol pathway enzymes can affect polyene sensitivity.

Research examining the sterol profiles of strains with modified ERG27 activity would be valuable for further elucidating its specific role in polyene resistance.

What is the relationship between ERG27 and echinocandin susceptibility in C. glabrata?

The relationship between ERG27 and echinocandin susceptibility is complex and likely indirect:

• Cell Wall-Membrane Interface: ERG27, through its role in ergosterol biosynthesis, affects membrane composition, which has downstream effects on cell wall organization and synthesis. Echinocandins target the cell wall by inhibiting β-(1,3)-D-glucan synthase .

• Stress Response Pathways: C. glabrata activates stress response pathways when exposed to antifungals. These pathways, including those regulated by transcription factors like Skn7p, Yap1p, Msn2p, and Msn4p, can be affected by alterations in membrane composition resulting from ERG27 dysfunction .

• Biofilm Formation: Proper ergosterol biosynthesis contributes to biofilm formation capacity, and biofilms provide protection against echinocandins. ERG27's role in maintaining ergosterol levels may therefore indirectly affect echinocandin efficacy in biofilm-forming populations .

While no direct evidence links ERG27 mutations to echinocandin resistance, the interconnection between membrane integrity (influenced by ERG27) and cell wall synthesis (targeted by echinocandins) suggests potential indirect effects. Further research specifically examining how ERG27 modifications affect echinocandin susceptibility would be valuable.

How does ERG27 interact with oxidosqualene cyclase (OSC/Erg7p) at the molecular level?

The interaction between ERG27 and OSC/Erg7p represents a fascinating example of protein-protein functional dependence, though the precise molecular details remain partially elusive:

• Functional Dependence: Research clearly demonstrates that yeast cells lacking ERG27 show concomitant loss of OSC activity, indicating that ERG27 is required for OSC functionality beyond its catalytic role as a steroid-3-ketoreductase .

• Separable Functions: Notably, a catalytically inactive ERG27 protein can still maintain its OSC protective function, suggesting that different domains within ERG27 mediate its catalytic activity versus its protein-protein interaction with OSC .

• Proposed Interaction Mechanisms:

  • Direct physical interaction, possibly involving chaperone-like activity

  • Co-localization in the endoplasmic reticulum membrane

  • Formation of a multi-enzyme complex within the ergosterol biosynthesis pathway

  • Structural stabilization of OSC through specific binding interfaces

• Species Differences: The requirement for ERG27 in OSC function appears to be specific to fungi. In mammalian cells, the ERG27 homolog (HSD17B7) is not required for OSC activity, highlighting an evolutionary divergence in sterol biosynthesis regulation .

The molecular basis for this interaction remains a significant research question. Techniques such as co-immunoprecipitation, yeast two-hybrid analysis, and cryo-electron microscopy of purified complexes could provide further insights into the structural basis of this functional relationship.

What role does ERG27 play in hypoxic adaptation in C. glabrata?

ERG27 contributes to hypoxic adaptation in C. glabrata through its integration with oxygen-dependent sterol biosynthesis regulation:

• Oxygen-Dependent Pathway: Ergosterol biosynthesis requires oxygen for several steps. Under hypoxic conditions, the pathway must be carefully regulated to adapt to limited oxygen availability .

• Transcriptional Control: In C. glabrata, transcription factors Zcf4 and Zcf27 (homologs of S. cerevisiae Hap1) regulate ERG genes under different oxygen conditions. Specifically, Zcf4 protein expression is induced under hypoxic conditions and represses ERG genes, including potentially ERG27 .

• Metabolic Adaptation: By modulating ergosterol pathway activity under low oxygen, C. glabrata can reallocate limited oxygen to essential processes while maintaining sufficient membrane function with altered sterol composition.

• Connection to Virulence: This hypoxic adaptation mechanism is particularly important in host environments where oxygen availability may be limited, such as in certain infection sites or within phagocytic cells .

The differential regulation of ergosterol pathway genes, including ERG27, under hypoxic conditions represents an important adaptation mechanism that allows C. glabrata to survive in diverse host environments. The specific binding of transcription factors like Zcf4 to ERG gene promoters under hypoxia has been demonstrated through chromatin immunoprecipitation studies .

How is ERG27 expression regulated in response to antifungal stress and environmental conditions?

ERG27 expression regulation in C. glabrata involves sophisticated responses to various stressors:

• Azole Response Regulation:

  • Zinc cluster transcription factors, particularly Zcf27, facilitate azole-mediated activation of ERG genes including ERG27

  • Treatment with azoles leads to increased binding of these transcription factors to ERG gene promoters

  • This upregulation helps compensate for inhibition of Erg11 (the primary azole target)

• Oxygen-Dependent Regulation:

  • Under hypoxic conditions, Zcf4 becomes specifically induced and represses ERG genes

  • This adaptation helps C. glabrata conserve oxygen usage when availability is limited

• General Stress Response Integration:

  • Stress-activated protein kinase (SAPK) Hog1 and transcription factors like Cap1 regulate stress pathways that may influence ERG27 expression

  • These factors respond to osmotic, oxidative, and nitrosative stress

  • DNA damage checkpoint kinase Rad53 may also play a role in this regulation

• Host Environment Adaptation:

  • During phagocytosis by host immune cells, C. glabrata activates stress response genes that may include modulation of ERG27 to adapt to the phagosomal environment

  • Factors like Skn7p, Yap1p, Msn2p, and Msn4p are involved in this adaptation

The regulation of ERG27 represents a critical aspect of C. glabrata's ability to adapt to diverse environmental conditions and stressors, including antifungal therapy, host immune responses, and varying oxygen availability.

How might structure-based drug design be applied to target C. glabrata ERG27?

Structure-based drug design targeting C. glabrata ERG27 represents a promising approach for novel antifungal development:

• Structural Analysis Prerequisites:

  • Determination of crystal structure of C. glabrata ERG27 through X-ray crystallography or cryo-EM

  • In silico modeling based on homologous proteins if experimental structures are unavailable

  • Identification of catalytic site and protein-protein interaction domains

• Target Site Selection:

  • Primary catalytic site: targeting the 3-ketosteroid reduction function

  • Secondary target: the OSC-interaction domain, which could disrupt the protein's moonlighting function

  • Allosteric sites that could affect either or both functions

• Selective Inhibition Strategy:

  • Design compounds that selectively inhibit fungal ERG27 without affecting human homologs (HSD17B7)

  • Exploit structural differences in substrate binding pockets between fungal and human enzymes

  • Target fungal-specific protein-protein interaction domains

• Methodological Approach:

  • Virtual screening of compound libraries against modeled binding sites

  • Fragment-based drug design to identify initial scaffolds

  • Structure-activity relationship studies to optimize lead compounds

  • Validation through enzymatic assays and whole-cell testing

The development of recombinant expression systems for C. glabrata HMGR has demonstrated the feasibility of producing active ergosterol pathway enzymes for inhibitor studies . Similar approaches with ERG27 could facilitate high-throughput screening for inhibitors.

What are the knowledge gaps in understanding ERG27's dual functionality?

Several significant knowledge gaps remain regarding ERG27's dual functionality:

• Structural Basis of Moonlighting Function:

  • The specific domains or residues responsible for OSC interaction versus catalytic activity remain incompletely characterized

  • Structural studies are needed to understand how one protein performs these distinct functions

  • It's unclear whether conformational changes mediate switching between functions

• Regulation of Dual Activities:

  • The mechanisms that regulate the balance between ERG27's catalytic and protein-interaction functions are poorly understood

  • Whether post-translational modifications control this balance remains unknown

  • The specific cellular conditions that might favor one function over the other need clarification

• Evolutionary Development:

  • How and why this dual function evolved in fungi but not mammals requires further investigation

  • Whether this represents an adaptation to specific fungal ecological niches or stressors

  • The evolutionary relationships between ERG27 and related enzymes across fungal species

• Interaction Network Complexity:

  • The complete set of proteins that interact with ERG27 beyond OSC/Erg7p remains to be fully characterized

  • Whether ERG27 forms part of a larger multi-enzyme complex in the ergosterol pathway

  • The spatial organization of these interactions within the endoplasmic reticulum

Addressing these gaps would require interdisciplinary approaches combining structural biology, biochemistry, genetics, and evolutionary analysis. Such research could reveal fundamental principles about protein moonlighting functions while also identifying novel antifungal targets.

How might ERG27 function contribute to C. glabrata pathogenesis in immunocompromised hosts?

ERG27's potential contributions to C. glabrata pathogenesis in immunocompromised hosts involve several interconnected mechanisms:

• Adaptation to Host Microenvironments:

  • ERG27's role in ergosterol biosynthesis helps C. glabrata adapt to varying oxygen levels in different host tissues

  • This adaptation is particularly important in immunocompromised hosts where fungi may access normally restricted tissue sites

  • The regulation by Zcf27 and Zcf4 allows fine-tuning of sterol composition based on environmental conditions

• Antifungal Treatment Survival:

  • ERG27's integration with azole response mechanisms may contribute to treatment failures in immunocompromised patients

  • The compensatory upregulation of ergosterol pathway components when Erg11 is inhibited by azoles can maintain membrane integrity

  • This contributes to both innate and acquired drug resistance

• Stress Response Integration:

  • C. glabrata can survive within phagocytes by generating strong stress responses against reactive oxygen species (ROS)

  • Proper ergosterol biosynthesis, dependent on ERG27, is critical for membrane integrity during stress responses

  • The activation of stress-response genes (Skn7p, Yap1p, Msn2p, Msn4p) helps neutralize the phagocytic environment

• Virulence Factor Support:

  • Proper membrane composition influences the expression and function of virulence factors like adhesins, proteases, and phospholipases

  • ERG27 indirectly contributes to biofilm formation capacity, which protects against host defenses and antifungals

  • In immunocompromised hosts with dysfunctional phagocytes, these adaptations may be particularly advantageous

Future research directions should include in vivo studies using ERG27 mutants in immunocompromised animal models to directly assess its contribution to virulence and pathogenesis.

How do mammalian homologs of ERG27 (HSD17B7) differ functionally from the fungal enzyme?

The functional divergence between fungal ERG27 and its mammalian homolog HSD17B7 reveals important evolutionary adaptations:

This functional divergence suggests that while the catalytic function is conserved, the protein-protein interaction function represents a fungal-specific adaptation. Mammalian HSD17B7 maintains catalytic activity when expressed in yeast but cannot complement the OSC-protective function of ERG27 . This fundamental difference may offer opportunities for selective targeting of fungal ERG27.

What methodological approaches are most effective for comparing ERG27 function across Candida species?

Effective methodological approaches for cross-species ERG27 functional comparison include:

• Complementation Studies:

  • Expression of ERG27 from different Candida species in C. glabrata ERG27-deletion mutants

  • Assessment of growth restoration on various media

  • Measurement of ergosterol production and OSC activity restoration

  • Analysis of antifungal susceptibility profiles

• Biochemical Characterization:

  • Heterologous expression and purification of ERG27 from multiple species

  • Comparative enzyme kinetics (Km, Vmax, substrate specificity)

  • Inhibition profiles using various compounds

  • Protein stability and cofactor requirements

• Structural Analysis:

  • Homology modeling based on conserved domains

  • Where possible, direct structural determination through crystallography or cryo-EM

  • Identification of species-specific structural features

  • In silico docking studies with substrates and inhibitors

• Evolutionary Analysis:

  • Phylogenetic analysis of ERG27 sequences across Candida species

  • Identification of conserved vs. variable regions

  • Selection pressure analysis to identify functionally important residues

  • Correlation of sequence variations with phenotypic differences

• Protein Interaction Studies:

  • Yeast two-hybrid or co-immunoprecipitation to compare interaction partners

  • Cross-species expression to identify compatibility of interaction interfaces

  • Localization studies to determine subcellular distribution patterns

These comparative approaches would provide valuable insights into species-specific adaptations in ERG27 function and potentially identify vulnerabilities that could be exploited for species-targeted antifungal development.

What experimental evidence supports the dual functionality of ERG27 in C. glabrata?

Substantial experimental evidence supports ERG27's dual functionality in C. glabrata:

• Genetic Evidence:

  • ERG27 deletion strains show loss of both 3-ketosteroid reductase activity and OSC function

  • The dual phenotype demonstrates that both functions depend on ERG27's presence

• Functional Separation Studies:

  • Catalytically inactive ERG27 mutants retain OSC protective function

  • This demonstrates that the two functions can be genetically separated

  • Suggests distinct structural domains responsible for each function

• Cross-Species Complementation:

  • Mammalian HSD17B7 expressed in ERG27-deficient yeast shows:

    • Detectable 3-ketoreductase activity

    • Almost complete absence of OSC activity restoration

    • Poor growth on sterol-deficient media

  • These results confirm that catalytic activity can be provided by homologs, but the OSC protection function is fungal-specific

• Biochemical Evidence:

  • Detection of accumulated 3-ketosteroids in strains with non-functional ERG27

  • Analytical methods (TLC, GC-MS) confirm the identity of these intermediates

  • NaBH4 treatment confirms the presence of 3-keto groups in accumulated intermediates

• Pathway Analysis:

  • Transcriptional regulation studies show that ERG27 is co-regulated with other ergosterol pathway genes

  • The gene is responsive to both azole stress and hypoxia

  • This integration in regulatory networks supports its central role in pathway function

This body of evidence collectively establishes that ERG27 performs dual functions that are mechanistically distinct yet both essential for proper ergosterol biosynthesis in C. glabrata.

What are the most promising therapeutic applications targeting ERG27 in antifungal drug development?

Based on current understanding, several promising therapeutic approaches targeting ERG27 emerge:

• Dual-Function Inhibitors:

  • Compounds targeting both the catalytic site and protein-interaction domains could provide synergistic effects

  • Such inhibitors would simultaneously block ergosterol synthesis and destabilize OSC

  • This dual mechanism could reduce the likelihood of resistance development

• Selective Targeting Based on Fungal-Specific Features:

  • Exploiting the structural differences between fungal ERG27 and mammalian HSD17B7

  • Focusing on the OSC-interaction domain, which appears to be fungal-specific

  • Developing compounds that selectively bind fungal ERG27 without affecting human homologs

• Combination Therapies:

  • Using ERG27 inhibitors alongside azoles to prevent compensatory upregulation

  • This approach could restore sensitivity in resistant strains

  • The complementary mechanisms of action could allow lower doses of each drug

• Species-Specific Targeting:

  • Developing inhibitors that exploit unique features of C. glabrata ERG27

  • This approach could provide options for targeted therapy based on species identification

  • Particularly valuable for treating multidrug-resistant C. glabrata infections

The recombinant expression systems developed for C. glabrata enzymes provide platforms for high-throughput screening of potential inhibitors. Future drug development should prioritize compounds that maintain activity against resistant isolates while exhibiting minimal effects on mammalian homologs.

What critical knowledge gaps remain to be addressed in ERG27 research?

Despite significant progress, several critical knowledge gaps persist:

• Structural Characterization:

  • No high-resolution structure of C. glabrata ERG27 is currently available

  • The structural basis for dual functionality remains theoretical

  • Understanding of the specific binding interface with OSC is limited

• Regulatory Networks:

  • The complete transcriptional and post-translational regulatory mechanisms controlling ERG27 expression and activity remain partially characterized

  • Integration with broader stress response pathways needs further elucidation

  • The specific roles of zinc cluster transcription factors in different conditions require further study

• Clinical Relevance:

  • Direct evidence linking ERG27 mutations to clinical antifungal resistance is limited

  • The prevalence of ERG27 variants in clinical isolates is poorly documented

  • The impact of host factors on ERG27 function during infection remains understudied

• Methodological Limitations:

  • Challenges in expressing and purifying full-length, active ERG27 with membrane domains

  • Limited availability of specific inhibitors for mechanistic studies

  • Difficulties in studying protein-protein interactions in membrane-associated complexes

• Therapeutic Development:

  • The druggability of ERG27 remains theoretical without validated inhibitors

  • Optimization parameters for selective targeting are not well-established

  • Potential off-target effects on human steroid metabolism require investigation

Addressing these gaps would require interdisciplinary approaches combining structural biology, genetics, biochemistry, and clinical research to fully elucidate ERG27's function and therapeutic potential.

How might emerging technologies advance our understanding of ERG27 function and regulation?

Emerging technologies offer promising approaches to address current limitations in ERG27 research:

• Advanced Structural Biology Techniques:

  • Cryo-electron microscopy for membrane protein complexes could reveal ERG27-OSC interactions

  • AlphaFold2 and other AI-based structure prediction methods may provide insights into regions difficult to characterize experimentally

  • Hydrogen-deuterium exchange mass spectrometry (HDX-MS) could identify dynamic binding interfaces

• CRISPR-Based Approaches:

  • CRISPR interference (CRISPRi) for tunable gene repression to study ERG27 dosage effects

  • CRISPR activation (CRISPRa) to upregulate ERG27 and assess effects on drug resistance

  • Base editing for precise introduction of point mutations to identify critical residues

• Single-Cell Technologies:

  • Single-cell transcriptomics to examine heterogeneity in ERG27 expression within populations

  • Microfluidic approaches to study ERG27 regulation under rapidly changing conditions

  • Live-cell imaging with fluorescent reporters to track dynamic regulation

• Chemogenomic and Proteomic Approaches:

  • Activity-based protein profiling to identify ERG27 interaction partners

  • Thermal proteome profiling to study drug binding and pathway effects

  • Chemogenomic profiling to identify synthetic lethal interactions with ERG27

• Advanced Computational Methods:

  • Molecular dynamics simulations to study conformational changes during catalysis and protein interaction

  • Systems biology approaches to integrate ERG27 function within broader metabolic networks

  • Machine learning for prediction of inhibitor binding and resistance mechanisms