BRK1 Human

What is Human BRK1 and what are its basic structural characteristics?

Human BRK1 (BRICK1, SCAR/WAVE actin-nucleating complex subunit) is a protein involved in the regulation of actin and microtubule organization. It functions as part of the WAVE complex that activates the Arp2/3 complex, which is essential for actin nucleation. The canonical human BRK1 protein consists of 75 amino acid residues with a molecular mass of approximately 8.7 kDa . The protein is predominantly localized in the cytoplasm and belongs to the BRK1 protein family. There are up to two different isoforms reported for this protein in humans . The protein's amino acid sequence, particularly when expressed recombinantly with histidine tags, can be extended to approximately 98 amino acids with a corresponding increase in molecular mass to around 11 kDa .

What are the common synonyms and identifiers for Human BRK1 in research databases?

When conducting literature searches or database queries on Human BRK1, researchers should be aware of various synonyms and identifiers. The protein is officially known as BRICK1, SCAR/WAVE actin-nucleating complex subunit, but it is also referred to by numerous alternative names including: C3orf10 (chromosome 3 open reading frame 10), haematopoietic stem/progenitor cell protein 300 (HSPC300), MDS027, hHBrk1, and simply BRICK1 . For database searches, the following identifiers are valuable: Gene ID: 55845, mRNA RefSeq: NM_018462, Protein RefSeq: NP_060932, MIM: 611183, UniProt ID: Q8WUW1. The gene is located on chromosome 3p25.3 . Using these various identifiers ensures comprehensive retrieval of research data across different biological databases when conducting literature reviews or comparative analyses.

What cellular pathways involve Human BRK1 and why are they significant?

Human BRK1 participates in several critical cellular signaling and structural pathways that influence cell morphology, motility, and response to environmental cues. Key pathways involving BRK1 include: ErbB1 downstream signaling, RAC1 signaling pathway, and regulation of actin cytoskeleton . These pathways are significant because they control fundamental cellular processes such as migration, adhesion, and cytoskeletal remodeling. The ErbB1 pathway influences cell proliferation and survival, while RAC1 signaling regulates cell movement and morphological changes. BRK1's role in actin cytoskeleton regulation is particularly crucial for cellular processes requiring coordinated membrane protrusion and retraction, such as cell migration during development, immune response, and wound healing. Understanding these pathway interactions provides insights into both normal cellular physiology and pathological conditions where cytoskeletal dysregulation occurs.

What are the optimal methods for detecting Human BRK1 in experimental samples?

For effective detection of Human BRK1 in experimental samples, researchers can employ several complementary techniques. Immunodetection methods are particularly valuable, with Western blotting (WB) serving as a primary approach for quantitative analysis of BRK1 expression levels. Immunohistochemistry (IHC) in both frozen (IHC-fr) and paraffin-embedded (IHC-p) tissue sections allows for visualization of BRK1's spatial distribution within tissues . Immunofluorescence (IF) provides higher resolution insights into subcellular localization, especially when combined with markers for other cytoskeletal components. Enzyme-linked immunosorbent assay (ELISA) offers quantitative measurement of BRK1 in solution . For optimal results, researchers should select antibodies with validated specificity for human BRK1, considering cross-reactivity with other species if performing comparative studies. Conjugated antibodies (e.g., with HRP or fluorophores like Alexa Fluor 594) can enhance detection sensitivity and enable multiplexed analyses when studying BRK1 alongside other proteins in the WAVE complex .

How can researchers effectively express and purify recombinant Human BRK1 for in vitro studies?

For successful expression and purification of recombinant Human BRK1, researchers should employ bacterial expression systems, particularly E. coli, which has been demonstrated as an effective host . The protein should be expressed with affinity tags to facilitate purification, with histidine tags (His-tag) being the most commonly utilized option. A typical construct would include the full 1-75 amino acid sequence of Human BRK1 fused to an N-terminal His-tag. Following expression, purification can be achieved through conventional chromatography techniques, particularly immobilized metal affinity chromatography (IMAC) leveraging the histidine tag's affinity for metal ions . For optimal stability and functionality, the purified protein should be stored in an appropriate buffer such as 20mM Tris-HCl (pH 8.0) containing 0.15M NaCl, 20% glycerol, and 1mM DTT as a reducing agent to maintain protein integrity . Researchers should verify protein purity (>90%) using SDS-PAGE and confirm the molecular mass using techniques such as MALDI-TOF mass spectrometry to ensure the expected mass of approximately 11 kDa for the His-tagged construct .

What IRB considerations apply when conducting Human BRK1 research using human tissue samples?

When conducting Human BRK1 research using human tissue samples, researchers must navigate specific Institutional Review Board (IRB) considerations. All projects that meet the definition of research and involve human subjects must undergo IRB review prior to any data collection to determine the appropriate level of review . There are three major types of IRB review relevant to BRK1 research: Exempt, Expedited, and Full review. BRK1 studies frequently fall under expedited review categories, particularly when they involve "collection of data through noninvasive procedures routinely employed in clinical practice" or "research involving materials (data, documents, records, or specimens) that have been collected, or will be collected, solely for non-research purposes" . For studies examining BRK1 in relation to individual or group characteristics or behavior, researchers must ensure that subject identities are protected. This is particularly important when the research involves "perception, cognition, motivation, identity, language, communication, cultural beliefs or practices, and social behavior" aspects that might intersect with BRK1's role in cellular function .

What statistical approaches are appropriate for validating Human BRK1 interaction data?

Validating Human BRK1 interaction data requires robust statistical approaches that account for both direct physical interactions and functional associations within the WAVE complex. For protein-protein interaction studies, researchers should implement statistical significance tests that account for the probability of random associations, particularly when using high-throughput techniques like affinity purification-mass spectrometry or yeast two-hybrid screens. Binary classification metrics such as precision-recall curves and receiver operating characteristic (ROC) analysis provide quantitative assessments of interaction prediction quality . When analyzing contradictory interaction data, researchers should apply Boolean minimization techniques to reduce complex interdependencies to their essential relationships, as the minimum number of Boolean rules (θ) required to assess contradictions is often significantly lower than the number of described contradictions (β) . For network-based analyses of BRK1's role in actin cytoskeleton regulation, statistical enrichment methods should be employed to identify significantly overrepresented pathways or functions. Bayesian approaches can be particularly valuable for integrating diverse data types (genomic, proteomic, functional) to build comprehensive models of BRK1's interaction landscape with appropriate confidence metrics.

How should researchers approach dimensional reduction for complex Human BRK1 datasets?

When analyzing complex multidimensional datasets related to Human BRK1, dimensional reduction techniques are essential for meaningful interpretation. For BRK1 research, which often generates high-dimensional data from proteomics, genomics, and cellular imaging, researchers should first assess the interdependency structure by identifying the number of interrelated variables (α) and their relationships . Principal Component Analysis (PCA) provides an initial approach for exploratory analysis, revealing major patterns of variation in BRK1 expression or activity across conditions. For datasets with nonlinear relationships, t-Distributed Stochastic Neighbor Embedding (t-SNE) or Uniform Manifold Approximation and Projection (UMAP) may better preserve local relationship structures important in understanding BRK1's context-dependent functions within the WAVE complex. When integrating heterogeneous data types, researchers should consider tensor decomposition methods that maintain the structural relationships between different data modalities. For contradiction analysis in multidimensional BRK1 datasets, Boolean minimization approaches can reduce complexity by determining the minimal number of rules (θ) required to characterize the contradictory relationships, allowing for more effective data quality assessment and biological interpretation .

How does Human BRK1 contribute to the regulation of actin and microtubule organization at the molecular level?

Human BRK1 serves as a critical regulatory component of the SCAR/WAVE actin-nucleating complex, functioning as an essential bridge that maintains the stability and proper assembly of this multiprotein complex . At the molecular level, BRK1 contributes to cytoskeletal organization through multiple mechanisms. It facilitates the activation of the Arp2/3 complex, which directly nucleates branched actin filament networks at the leading edge of migrating cells . This process is crucial for lamellipodium formation and cell motility. BRK1's function extends beyond actin regulation to include microtubule organization, suggesting an integrated role in coordinating the entire cytoskeletal network . The protein's small size (8.7 kDa) belies its structural importance as it helps maintain the integrity of the much larger WAVE complex. Research indicates that BRK1 likely functions through protein-protein interactions mediated by specific structural domains within its 75-amino-acid sequence . These interactions enable signal transduction from upstream regulators like Rac1 to the actin polymerization machinery, effectively translating extracellular cues into cytoskeletal rearrangements that drive cellular behaviors including migration, adhesion, and morphogenesis.

What are the implications of Human BRK1 in cellular pathologies and disease mechanisms?

The involvement of Human BRK1 in fundamental cytoskeletal regulatory processes suggests significant implications for various cellular pathologies and disease mechanisms. As a component of the WAVE complex that regulates actin dynamics, BRK1 dysfunction may contribute to diseases characterized by aberrant cell migration, adhesion, or morphology . Given its role in ErbB1 downstream signaling pathways, alterations in BRK1 function could potentially impact cancer progression, particularly in processes requiring cytoskeletal remodeling such as invasion and metastasis . The protein's involvement in hematopoietic stem/progenitor cells (as suggested by its synonym HSPC300) indicates potential roles in hematological disorders if dysregulated . The location of the BRK1 gene on chromosome 3p25.3 places it in a region associated with various genetic disorders and susceptibility loci . While direct clinical evidence linking BRK1 mutations to specific diseases remains limited in the provided search results, its fundamental role in cellular architecture suggests that even subtle alterations could have cascading effects on cell function. Research methodologies focusing on BRK1 in pathological contexts should combine genetic analyses with functional studies of cytoskeletal dynamics to elucidate how BRK1 variants might contribute to disease phenotypes.

What cutting-edge techniques are advancing our understanding of Human BRK1 dynamics and interactions?

Cutting-edge techniques are rapidly advancing our understanding of Human BRK1 dynamics and interactions within the complex cellular environment. Super-resolution microscopy techniques, including Structured Illumination Microscopy (SIM), Stimulated Emission Depletion (STED), and Single-Molecule Localization Microscopy (SMLM), now allow visualization of BRK1 localization and dynamics at nanometer resolution, revealing previously unobservable details of its spatial organization within the WAVE complex . Proximity labeling approaches such as BioID and APEX2 are enabling the mapping of BRK1's protein interaction neighborhood in living cells, providing temporal information about dynamic interaction partners during cytoskeletal remodeling events. For structural studies, cryo-electron microscopy is offering insights into the three-dimensional conformation of BRK1 within the assembled WAVE complex. Techniques for studying protein dynamics, including Förster Resonance Energy Transfer (FRET) and Fluorescence Recovery After Photobleaching (FRAP), are illuminating the kinetics of BRK1 incorporation into the WAVE complex. At the genetic level, CRISPR-Cas9 genome editing is enabling precise manipulation of BRK1 to create cellular models for functional studies . Integration of these techniques with computational approaches for contradiction analysis and data validation is essential for building consistent models of BRK1 function across different experimental paradigms .

What are the most effective antibodies and detection reagents for Human BRK1 research?

For Human BRK1 research, selecting the most effective antibodies and detection reagents is crucial for experimental success. Based on validated research applications, polyclonal antibodies raised in rabbits have demonstrated reliable detection of Human BRK1 in multiple applications including Western blot (WB), immunohistochemistry (IHC), and immunofluorescence (IF) . When selecting antibodies, researchers should prioritize those validated specifically for Human BRK1 (also known as C3orf10) with demonstrated reactivity to human samples. For cross-species studies, antibodies with verified reactivity to both human and mouse BRK1 are available and allow for comparative analyses . For enhanced detection sensitivity, researchers can utilize antibodies conjugated to horse radish peroxidase (HRP) for chemiluminescent applications or fluorophores such as Alexa Fluor 594 for fluorescence microscopy . The chosen detection method should align with the experimental question, with IF providing superior resolution for subcellular localization studies, while WB offers quantitative assessment of expression levels. Researchers should be aware that antibody concentrations typically range from 100 μL to 100 μg per application, and validation data including positive and negative controls should be thoroughly reviewed before selection .

How can researchers design experiments to study the role of Human BRK1 in specific cellular pathways?

Designing experiments to study Human BRK1's role in specific cellular pathways requires a multifaceted approach that combines genetic manipulation, protein interaction studies, and functional assays. Researchers should begin with loss-of-function studies using RNA interference (siRNA/shRNA) or CRISPR-Cas9 genome editing to deplete BRK1 and observe the effects on pathway activity. For gain-of-function approaches, overexpression of tagged BRK1 constructs (with tags such as His or Myc/DDK) can help identify dose-dependent effects and enable co-immunoprecipitation studies to map pathway-specific interaction partners . When examining BRK1's role in actin dynamics specifically, researchers should incorporate live-cell imaging with fluorescently labeled actin to visualize cytoskeletal changes in real-time. For pathway-specific analyses, consider using pharmacological inhibitors of key nodes (such as RAC1 inhibitors when studying the RAC1 signaling pathway) in combination with BRK1 manipulation to identify epistatic relationships . To study BRK1's role in the ErbB1 downstream signaling pathway, stimulation with the appropriate ligands followed by biochemical assessment of pathway activation markers would be appropriate. For all experimental designs, researchers must implement appropriate data quality controls and contradiction analysis methods to ensure the reliability of the results, especially when integrating multi-omics data from different experimental platforms .

What bioinformatics tools are essential for analyzing Human BRK1 sequence, structure, and functional relationships?

Bioinformatic analysis of Human BRK1 requires specialized tools that address its unique characteristics as a small but functionally significant protein. For sequence analysis, BLAST remains fundamental for identifying homologs across species, but researchers should apply position-specific scoring matrices to detect distant relationships given BRK1's relatively short 75-amino acid sequence . Structural prediction tools including AlphaFold2 can generate high-confidence models of BRK1's three-dimensional conformation, particularly valuable given the challenges of crystallizing small proteins within larger complexes. For functional analysis, researchers should utilize Gene Ontology enrichment tools to contextualize BRK1 within biological processes, molecular functions, and cellular components, with particular attention to cytoskeletal organization terms . Protein-protein interaction databases such as STRING and BioGRID provide essential information about BRK1's interactome, while pathway analysis tools like KEGG and Reactome offer insights into its role in signaling cascades including ErbB1 downstream and RAC1 pathways . For contradictions in multi-omics datasets involving BRK1, researchers should implement R-based data quality assessment packages, though awareness of their limitations for complex contradiction patterns beyond the (2,1,1) class is important . Evolutionary analysis tools like PAML can provide insights into selective pressures on BRK1 across species, offering clues about functionally important regions conserved through evolution.

What are the current knowledge gaps in Human BRK1 research that present opportunities for novel investigations?

Current knowledge gaps in Human BRK1 research present several compelling opportunities for novel investigations. Despite understanding BRK1's general role in the WAVE complex and actin nucleation, the precise molecular mechanisms by which this small 8.7 kDa protein exerts its regulatory effects on the substantially larger WAVE complex remains incompletely characterized . The reported existence of up to two isoforms of Human BRK1 raises questions about their potentially distinct functions in different cellular contexts or developmental stages that have yet to be thoroughly explored . While BRK1's involvement in cytoskeletal regulation is established, its specific contributions to microtubule organization represent an under-investigated area that could reveal important cytoskeletal crosstalk mechanisms . The protein's participation in multiple signaling pathways (ErbB1 downstream, RAC1, actin cytoskeleton regulation) suggests integration points between these pathways that warrant systematic investigation . Additionally, the potential connections between BRK1 function and human diseases remain largely unexplored, particularly in conditions characterized by cytoskeletal dysregulation. The development of methodologies for addressing data contradictions in complex BRK1 interaction networks presents both a challenge and an opportunity to improve research quality and reproducibility .

How might emerging technologies advance Human BRK1 research in the next decade?

Emerging technologies will likely transform Human BRK1 research in the next decade by enabling more precise, dynamic, and integrated analyses of this important cytoskeletal regulator. Spatially-resolved transcriptomics and proteomics will allow researchers to map BRK1 expression and activity within specific cellular microenvironments and tissue architectures, providing contextual information about its function. Advanced cryo-electron tomography techniques will likely reveal the molecular details of BRK1 within the intact WAVE complex at near-atomic resolution, illuminating its structural contributions. Optogenetic and chemogenetic tools will enable temporal control of BRK1 activity, allowing researchers to dissect its acute roles in cytoskeletal dynamics with unprecedented precision . Single-cell multi-omics approaches will uncover cell-type specific functions and regulatory networks involving BRK1, particularly in heterogeneous tissues. Artificial intelligence and machine learning algorithms will accelerate the identification of patterns in complex datasets, potentially resolving contradictions in experimental findings through sophisticated Boolean minimization and pattern recognition . CRISPR-based lineage tracing combined with BRK1 functional manipulation will elucidate its developmental roles across cellular differentiation trajectories. These technological advances, coupled with improved computational frameworks for handling multidimensional data dependencies, will substantially enhance our understanding of BRK1's diverse functions in normal physiology and disease contexts .