ISOC2 Human

What is ISOC2 and what is its genomic location?

ISOC2 is a protein-coding gene that belongs to the isochorismatase family. It is located on chromosome 19 in humans and encodes a mitochondrial protein that has significant roles in cellular metabolism . The protein contains an isochorismatase domain, which is characteristic of enzymes involved in various metabolic pathways. ISOC2 has multiple cellular localizations, being found in the mitochondrion, cytoplasm, and nucleus, suggesting diverse functional roles depending on cellular context .

What diseases are associated with ISOC2?

ISOC2 has been implicated in several significant neurodegenerative and developmental disorders. Research has identified associations with:

• Parkinson disease

• Lysosomal storage disease

• Alzheimer disease

• Multiple sclerosis

• Craniofacial microsomia

The involvement of ISOC2 in these diverse pathologies suggests its critical role in cellular processes that, when dysregulated, contribute to disease pathogenesis. The mechanisms through which ISOC2 affects these conditions remain an active area of research, particularly in neurodegenerative disease models.

How does ISOC2 interact with other proteins, particularly CDKN2A?

ISOC2 has been shown to interact with CDKN2A (cyclin-dependent kinase inhibitor 2A), an important tumor suppressor protein. This interaction is particularly notable because it influences the subcellular localization of ISOC2. Research indicates that ISOC2 localizes to the nucleus specifically in the presence of CDKN2A . This interaction suggests ISOC2 may play roles in cell cycle regulation, cellular senescence, or tumor suppression pathways.

Methodologically, researchers investigating this interaction typically employ:

• Co-immunoprecipitation (Co-IP) assays

• Fluorescence microscopy with tagged proteins

• Proximity ligation assays (PLA)

• Yeast two-hybrid screening

These approaches allow for the characterization of the interaction dynamics between ISOC2 and CDKN2A under various cellular conditions and stress responses.

What are the expression patterns of ISOC2 across different tissues?

ISOC2 expression has been extensively profiled across tissues through resources like The Human Protein Atlas. The protein shows distinct expression patterns across various tissues and brain regions, which may correlate with its functional importance in specific cell types . According to the Ma'ayan Laboratory data, ISOC2 has 3,706 functional associations with biological entities spanning 8 categories, including tissue-specific expression patterns .

Research methodologies to analyze expression patterns include:

• RNA-sequencing

• Immunohistochemistry

• Microarray analysis

• Single-cell transcriptomics

The Allen Brain Atlas datasets reveal specific expression patterns in human brain tissues, with both developmental and adult expression profiles available . These datasets allow researchers to analyze temporal and spatial expression patterns of ISOC2 throughout human development and across brain regions.

What are the recommended protocols for storing and handling recombinant ISOC2?

For researchers working with recombinant ISOC2 protein, proper storage and handling are critical to maintain protein stability and functionality. The recommended protocols include:

• Short-term storage (2-4 weeks): Store at 4°C

• Long-term storage: Store frozen at -20°C

• Formulation: The ISOC2 solution (0.5mg/ml) should contain 20mM Tris-HCl buffer (pH 8.0), 0.4M Urea, and 10% glycerol

• Stability enhancement: For long-term storage, add a carrier protein (0.1% HSA or BSA)

• Avoid multiple freeze-thaw cycles as they can compromise protein integrity

These protocols are essential to maintain the structural and functional properties of the protein for experimental applications, particularly for enzymatic assays or protein-protein interaction studies.

What experimental approaches are most effective for studying ISOC2 function?

Studying ISOC2 function requires a multifaceted approach that integrates various methodologies:

Genetic Manipulation Technologies:

• CRISPR/Cas9 gene editing to create knockout or knockin models

• siRNA or shRNA for transient knockdown studies

• Overexpression systems with tagged proteins

Biochemical Characterization:

• Enzymatic activity assays focusing on isochorismatase function

• Protein-protein interaction mapping using mass spectrometry

• Structural studies (X-ray crystallography, cryo-EM) to elucidate functional domains

Cellular Localization Studies:

• Immunofluorescence microscopy with subcellular markers

• Subcellular fractionation followed by Western blotting

• Live-cell imaging with fluorescently tagged ISOC2

Systems Biology Approaches:

• Integration of transcriptomic, proteomic, and metabolomic data

• Network analysis of ISOC2 interactions using resources like Harmonizome

• Computational modeling of metabolic pathways involving ISOC2

These methodological approaches should be selected based on the specific research question and available resources, with consideration for the multiple cellular localizations of ISOC2.

How can researchers reconcile contradictory data regarding ISOC2 localization?

The reported localization of ISOC2 in mitochondria, cytoplasm, and nucleus presents challenges in understanding its function . To address contradictory localization data, researchers should:

• Employ multiple detection methods:

  • Antibody validation using multiple independent antibodies

  • Complementary approaches (subcellular fractionation, immunofluorescence, and electron microscopy)

  • Tagged protein variants with different tags at different positions

• Consider context-dependent localization:

  • Evaluate cell-type specific localization patterns

  • Assess localization under different physiological conditions

  • Investigate the role of CDKN2A in nuclear localization

• Temporal dynamics investigation:

  • Time-course experiments after stimulus

  • Cell-cycle dependent localization analysis

  • Developmental stage-specific localization patterns

• Isoform-specific analysis:

  • Determine if different protein isoforms have distinct localization patterns

  • Consider post-translational modifications affecting localization

By implementing these methodological approaches, researchers can develop a more nuanced understanding of ISOC2's dynamic localization patterns and reconcile apparently contradictory observations.

What methodological considerations are important for studying ISOC2 in disease models?

When investigating ISOC2 in disease models, particularly neurodegenerative conditions, several methodological considerations are critical:

Disease Model Selection:

• Cellular models (patient-derived iPSCs, neuronal cultures)

• Animal models (transgenic mice, Drosophila, C. elegans)

• Organoid systems for 3D tissue architecture

Disease-Relevant Assays:

• Mitochondrial function assessment (given ISOC2's mitochondrial localization)

• Protein aggregation studies for neurodegenerative conditions

• Lysosomal function assays for lysosomal storage diseases

Temporal Considerations:

• Age-dependent studies for late-onset diseases like Parkinson's and Alzheimer's

• Developmental timing for craniofacial microsomia research

• Disease progression monitoring in longitudinal studies

Translational Approaches:

• Correlation of findings between model systems and human tissues

• Biomarker development based on ISOC2 function or levels

• Therapeutic targeting strategies considering subcellular localization

When studying ISOC2 in disease contexts, researchers should incorporate relevant disease endpoints and consider the specific pathological mechanisms of each condition while maintaining rigorous controls for genetic background and environmental factors.

What are emerging areas of ISOC2 research with therapeutic potential?

Given ISOC2's associations with multiple neurodegenerative diseases, several promising research directions may yield therapeutic insights:

• Small molecule modulators of ISOC2 function:

  • High-throughput screening for isochorismatase activity modulators

  • Structure-based drug design targeting specific ISOC2 domains

  • Allosteric modulators of protein-protein interactions

• ISOC2 in mitochondrial dysfunction:

  • Investigate ISOC2's role in mitochondrial metabolism pathways

  • Explore connections to mitochondrial dysfunction in Parkinson's disease

  • Develop mitochondria-targeted ISOC2-based interventions

• ISOC2-CDKN2A axis in neurodegeneration:

  • Characterize the functional consequences of this interaction

  • Develop peptide inhibitors or enhancers of the interaction

  • Investigate cell cycle re-entry in post-mitotic neurons

These emerging research areas require innovative methodological approaches and cross-disciplinary collaboration between structural biologists, medicinal chemists, and neuroscientists to translate basic ISOC2 research into therapeutic applications.