MAK16 Human

What is the molecular identity of human MAK16?

MAK16 (also known as MAK16L or RBM13) is a homolog of the S. cerevisiae MAK16 gene that enables RNA binding activity . The human gene (ID: 84549) encodes a protein with significant evolutionary conservation across multiple species, including mouse (67920), rat (306526), and numerous other mammals . This high degree of conservation suggests critical biological functions. MAK16 interacts extensively with components of the 60S ribosomal protein, indicating its potential involvement in ribosome biogenesis or function .

How can researchers detect and analyze MAK16 expression?

Several validated research tools are available for MAK16 expression analysis:

Table 1: Research Tools for MAK16 Detection

For comprehensive protein-protein interaction studies, researchers should employ combinatorial approaches including co-immunoprecipitation followed by mass spectrometry, proximity labeling techniques (BioID/APEX), and yeast two-hybrid screening to validate MAK16's interactome .

What is the significance of MAK16 in autoimmune disease research?

Recent proteomic research has identified anti-MAK16 autoantibodies as novel biomarkers for systemic lupus erythematosus (SLE). In a comprehensive two-phase study involving 125 SLE patients, 111 healthy volunteers, and 60 disease controls, anti-MAK16 autoantibodies showed significantly higher expression levels in SLE patients compared to both control groups . This represents one of several previously unreported autoantibodies discovered through human proteome microarray screening technology.

What experimental methods effectively detect anti-MAK16 autoantibodies in clinical samples?

Researchers employing either proteome microarray or ELISA methodology can reliably detect anti-MAK16 autoantibodies:

ELISA Protocol for Anti-MAK16 Detection:

• Coat 96-well plates with purified MAK16 protein

• Add patient serum samples (dilution optimization required)

• Add horseradish peroxidase-labeled anti-human IgG

• Measure optical density values via microplate reader

The study validated consistent results between protein microarray and ELISA methods, confirming methodological reliability across platforms .

What is the diagnostic performance of anti-MAK16 for autoimmune disease?

Anti-MAK16 demonstrates clinically meaningful diagnostic potential for SLE:

Table 2: Diagnostic Performance of Anti-MAK16 in SLE

While anti-MAK16 alone shows moderate diagnostic potential, its optimal clinical utility was demonstrated in a multimarker panel. When combined with anti-RPLP2, anti-PARP1, and anti-RPL7A, the diagnostic model achieved superior performance with AUC values of 0.828-0.932 across different comparison groups .

How should researchers design loss-of-function experiments for MAK16?

When conducting MAK16 knockdown studies, multiple experimental approaches should be considered:

• siRNA approach: Utilize commercially available pre-designed siRNAs based on the Rosetta Inpharmatics algorithm that specifically target MAK16 mRNA .

• shRNA approach: For stable knockdown, researchers should select from validated shRNA collections, ideally testing 3-5 different constructs to identify optimal knockdown efficiency .

• CRISPR-Cas9 methodology: While not specifically mentioned in the search results, this approach represents the gold standard for complete gene knockout studies.

For all approaches, researchers must include appropriate controls (scrambled sequences, empty vectors) and validate knockdown efficiency at both mRNA (qRT-PCR) and protein (western blot) levels before assessing functional outcomes.

What experimental designs best elucidate MAK16's role in ribosome biogenesis?

Given MAK16's interaction with 60S ribosomal proteins , researchers investigating its role in ribosome biogenesis should employ:

• Ribosome profiling: To analyze how MAK16 affects ribosome assembly and function.

• Polysome profiling: To assess how MAK16 knockdown affects translational efficiency.

• rRNA processing assays: To determine if MAK16 participates in pre-rRNA processing steps.

• Nucleolar localization studies: Using the validated antibodies (HPA050574, HPA044417) to visualize MAK16's subcellular distribution.

These approaches should be conducted in both normal conditions and under ribosomal stress to fully characterize MAK16's dynamic functions.

How can AI systems assist in analyzing complex MAK16 experimental data?

AI integration presents significant opportunities for analyzing MAK16-related datasets, though current human-AI interaction frameworks remain limited by "simplistic collaboration paradigms" . Researchers should be aware that AI systems trained on biological data may inherit biases that influence interpretations of experimental results . To maximize AI utility while minimizing bias:

• Train models on diverse datasets spanning multiple cell types and experimental conditions

• Implement transparent algorithmic decision-making processes

• Design truly interactive AI systems that facilitate bidirectional collaboration rather than one-way recommendations

What experimental approaches can address potential confounding factors in MAK16 autoantibody research?

When studying MAK16 autoantibodies in SLE, researchers must address several methodological challenges:

• Cross-reactivity assessment: Test for epitope cross-reactivity with other ribosomal proteins given MAK16's interactions with 60S ribosomal components .

• Temporal dynamics: Design longitudinal studies to determine whether anti-MAK16 antibodies appear early in disease progression or correlate with disease activity.

• Cellular localization experiments: Investigate whether MAK16 relocalization occurs during apoptosis or cellular stress, potentially exposing normally sequestered epitopes.

• Epitope mapping: Identify specific MAK16 regions recognized by autoantibodies using peptide array technology or hydrogen-deuterium exchange mass spectrometry.

These approaches would help distinguish whether anti-MAK16 represents a causative factor in disease pathogenesis or a secondary phenomenon.

How can researchers investigate MAK16's evolutionary significance?

The extensive conservation of MAK16 across species provides opportunities for evolutionary biology investigations:

Table 3: Selected MAK16 Homologs Across Species

Researchers should perform comparative genomic analyses to identify both highly conserved domains (suggesting core functions) and divergent regions (indicating species-specific adaptations). Multi-species functional complementation studies would help determine the degree of functional conservation.

What methodologies can resolve contradictions in MAK16 functional data?

When encountering contradictory findings regarding MAK16 function, researchers should employ:

• Cell type-specific analyses: Assess MAK16 function across diverse cell types, as its role may vary between proliferating and differentiated cells.

• Conditional knockdown systems: Utilize inducible systems to distinguish between direct and compensatory effects of MAK16 depletion.

• Proteomic interaction maps under different conditions: Compare MAK16 interactomes under various cellular states (proliferation, stress, differentiation).

• Single-cell approaches: Implement single-cell RNA-seq and proteomics to detect cell-to-cell variability in MAK16 function that may explain seemingly contradictory population-level data.

These methodological approaches would help reconcile discrepant findings and build a more comprehensive understanding of MAK16's multifaceted biological roles.