Recombinant Mouse Coronin-2A (Coro2a)

What is the basic structure of mouse Coronin-2A protein?

Mouse Coronin-2A (Coro2a) is a type-II coronin protein characterized by a distinct structural organization. The protein contains an N-terminal β-propeller region with seven WD40 repeats, a unique region (U region), and a C-terminal coiled-coil region (CC) . The WD40 repeats form a β-propeller structure that serves as a platform for protein-protein interactions, particularly with actin filaments. The unique region provides specificity for particular binding partners, while the coiled-coil domain mediates dimerization and interactions with other proteins. This structural arrangement enables Coro2a to function as a versatile regulator of actin dynamics and other cellular processes.

What are the main subcellular localizations of Coronin-2A in mouse cells?

Coronin-2A demonstrates multiple distinct subcellular localizations that reflect its diverse functions. It primarily localizes to actin stress fibers, with approximately 80% overlap with Phalloidin-labeled actin structures . This localization is specific, as treatment with cytochalasin D (an actin polymerization inhibitor) disrupts this pattern . Coro2a also partially localizes to focal adhesions, particularly in the internal regions of cells . Additionally, it partially overlaps with EHD1 on endosomal structures, reflecting its role in endosomal trafficking and fission processes . Coronin-2A is also found diffusely in the cytoplasm . For visualization studies, researchers have successfully used FLAG-tagged and GFP-tagged Coro2a constructs to observe these localization patterns in live cells .

How does Coronin-2A regulate actin dynamics?

Coronin-2A functions as a negative regulator of actin branching, similar to other coronin family members. Depletion of Coro2a results in dramatically expanded actin-rich protrusions at the plasma membrane, with quantitative analysis showing that these protrusions increase both in size and frequency following Coro2a knockdown . This phenotype suggests that Coro2a normally attenuates actin branching, potentially by disassembling Arp2/3-containing branched actin filaments .

The regulation of actin dynamics by Coro2a involves coordination with other actin-regulatory proteins. Research has revealed a functional relationship between Coro2a and the Slingshot-cofilin pathway, where increased Slingshot activity can partially compensate for Coro2a depletion . This interaction network creates a sophisticated regulatory system for controlling actin filament organization and turnover, which has important implications for processes like cell migration and adhesion dynamics.

What is known about the relationship between Coro2a and focal adhesion turnover?

Coro2a plays a critical role in regulating focal adhesion dynamics, particularly affecting their disassembly. Studies show that depletion of Coronin-2A increases focal adhesion size while decreasing the total number of focal adhesions per cell . The most significant effect occurs during focal adhesion disassembly, where the rate decreased by approximately half in Coro2a-depleted cells, while assembly rates remained unaffected .

The table below summarizes the effects of Coro2a depletion on focal adhesion dynamics:

What functional domains of Coronin-2A are critical for its interaction with other proteins?

Several functional domains in Coronin-2A mediate its interactions with binding partners. The most notable is the NPFDD sequence motif (asparagine-proline-phenylalanine-aspartic acid-aspartic acid) that preferentially binds to EHD1, connecting Coro2a to the endosomal fission machinery . This interaction provides a molecular link between actin dynamics and membrane trafficking processes.

The WD40 repeats in the N-terminal β-propeller region facilitate interactions with actin filaments and possibly other cytoskeletal proteins . The C-terminal coiled-coil domain is involved in protein oligomerization and interactions with components of the nuclear receptor corepressor complex, where Coro2a serves as a key component essential for recruiting histone deacetylases to form repressive chromatin structures .

What are the optimal approaches for detecting and analyzing mouse Coronin-2A in experimental systems?

For detecting and analyzing mouse Coronin-2A, researchers can employ several validated approaches:

• Antibody-based detection: Coronin 2A Antibody (H-2) from Santa Cruz Biotechnology (sc-376194) has been validated for detecting Coro2a from mouse, rat, and human origins through multiple applications including western blotting, immunoprecipitation, immunofluorescence, immunohistochemistry with paraffin-embedded sections, and ELISA .

• Expression constructs: Both FLAG-tagged and GFP-tagged Coro2a constructs have been successfully used for localization studies . When designing these constructs, consider that human and mouse Coro2a show high conservation, allowing human constructs to be used for rescue experiments in mouse cells.

• Localization studies: For visualizing Coro2a localization, co-staining with Phalloidin effectively demonstrates its association with actin filaments . Treatment with cytochalasin D can serve as a control to disrupt actin structures and confirm specificity of localization.

• Protein interaction analysis: Co-immunoprecipitation approaches can identify binding partners, particularly focusing on interactions with EHD1 through the NPFDD motif and components of actin-regulatory pathways .

When analyzing Coro2a in experimental systems, it's crucial to include appropriate controls and consider potential tissue or cell-type specific variations in expression and function.

How can I effectively knockdown Coro2a expression in mouse cell lines?

Multiple validated approaches for depleting Coronin-2A have been described in the literature:

• siRNA knockdown: This approach has been successfully used to reduce Coro2a expression, with validation by immunoblotting . When designing siRNA experiments, include non-targeting controls and validate knockdown efficiency through both protein and mRNA quantification.

• shRNA via lentiviral delivery: Developing a shRNA against Coro2a delivered by lentivirus provides a more stable knockdown . This approach should include:

  • Non-specific shRNA (shNS) as a control

  • Verification by both immunoblotting and qRT-PCR analysis

  • A rescue construct encoding both the shRNA and an RNAi-resistant Coro2a variant (e.g., human Coro2a-GFP)

When evaluating Coro2a knockdown effects, it's important to consider the following experimental design elements:

• Use multiple independent siRNA/shRNA sequences to confirm specificity

• Include appropriate controls for each assay

• Document phenotypes through quantitative measurements

• Consider potential compensatory mechanisms by related proteins

The rescue experiments are particularly crucial to confirm that observed phenotypes are specifically due to Coro2a depletion rather than off-target effects .

What protocols are recommended for studying Coro2a's role in cell migration?

To investigate Coro2a's role in cell migration, several complementary approaches have been validated:

For all migration assays, it's important to optimize conditions for your specific cell type and include appropriate controls, including both negative controls and rescue experiments to confirm specificity.

How should I design experiments to study the role of Coro2a in endosomal trafficking?

Designing experiments to investigate Coro2a's role in endosomal trafficking requires multiple complementary approaches:

• Cargo trafficking assays:

  • Examine internalization of clathrin-dependent cargo (transferrin, EGF receptor)

  • Assess uptake of clathrin-independent cargo (MHC-1, CD98 receptors)

  • Measure recycling rates of different cargo types

  • Compare these processes between control and Coro2a-depleted cells

• Endosomal morphology analysis:

  • Use confocal microscopy to examine endosomal size and distribution

  • Quantify endosome enlargement in Coro2a-depleted cells

  • Co-stain with markers for different endosomal compartments

  • Apply 3D reconstruction techniques for detailed morphological analysis

• Protein interaction studies:

  • Investigate the interaction between Coro2a and EHD1 via the NPFDD motif

  • Perform co-immunoprecipitation or proximity ligation assays

  • Create mutants of the NPFDD motif to disrupt specific interactions

  • Examine co-localization on endosomal structures using high-resolution microscopy

The research has established that Coro2a depletion causes different effects on distinct cargo types, with marked decrease in internalization of clathrin-dependent cargo but little impact on clathrin-independent cargo uptake . These differential effects highlight the importance of examining multiple cargo types in your experimental design.

What controls should be included when studying Coro2a function in cytoskeletal organization?

When investigating Coro2a's function in cytoskeletal organization, include the following essential controls:

• Knockdown validation controls:

  • Non-targeting siRNA/shRNA control

  • Western blot and qRT-PCR to confirm knockdown efficiency

  • Rescue experiments with RNAi-resistant Coro2a constructs

• Actin organization controls:

  • Cytochalasin D treatment to disrupt actin filaments

  • Phalloidin staining to visualize F-actin structures

  • 3D reconstruction of actin filaments using specialized software like Imaris

• Quantification controls:

  • Automated, unbiased measurement of actin-rich protrusions

  • Analysis of filament tracer tool reconstructions

  • Quantification of the frequency and mean diameter of actin-rich protrusions

• Cell-type controls:

  • Compare effects across different cell types (e.g., U251 Glioblastoma cells show very elaborate actin cytoskeletons)

  • Consider analyzing both normal and transformed cell lines

• Functional assay controls:

  • For migration studies, include positive controls (e.g., cytochalasin D for complete inhibition)

  • For adhesion studies, use focal adhesion markers like paxillin (PXN)

  • Time-course experiments to track temporal changes in cytoskeletal organization

These comprehensive controls ensure that observed phenotypes are specifically attributable to Coro2a function and provide a foundation for accurate interpretation of experimental results.

How does the NPFDD sequence motif in Coro2a mediate its interaction with EHD1?

The NPFDD sequence motif in Coronin-2A represents a specialized binding domain that mediates a critical interaction with EHD1, a key component of the endosomal fission machinery . This motif consists of asparagine-proline-phenylalanine-aspartic acid-aspartic acid residues that create a specific recognition site for EH domains present in EHD1.

The molecular basis of this interaction involves the NPF portion of the motif, which is a known binding sequence for EH domains. The additional DD (aspartic acid-aspartic acid) residues provide additional specificity for EHD1 binding . This interaction facilitates the partial overlap of Coro2a with EHD1 on endosomal structures and establishes a functional connection between actin regulation and endosomal fission processes.

For researchers investigating this interaction, approaches include:

• Mutational analysis of the NPFDD motif to determine essential residues

• In vitro binding assays to characterize interaction affinity and specificity

• Structural studies of the EHD1-Coro2a complex

• Functional assays to determine how disruption of this interaction affects endosomal fission and recycling

This interaction represents a molecular bridge connecting the actin cytoskeleton to membrane trafficking machinery, highlighting one mechanism by which Coro2a coordinates diverse cellular processes .

What is the mechanistic relationship between Coro2a and the Slingshot-cofilin pathway?

Research has revealed a sophisticated functional relationship between Coronin-2A and the Slingshot-cofilin pathway in regulating actin dynamics and focal adhesion turnover. The evidence indicates that increased Slingshot activity can partially compensate for depletion of coronin 2A .

Mechanistically, this relationship involves several components:

• Slingshot-1L (SSH1L) is a phosphatase that activates cofilin by dephosphorylation

• Cofilin promotes actin filament disassembly and turnover

• Coronin-2A appears to work in concert with this pathway to regulate focal adhesion disassembly

Experimental evidence shows that expression of a constitutively active Slingshot-1L mutant (SSH1L-2SA) partially rescued the focal adhesion disassembly defect in coronin-2A-depleted cells . The rate of disassembly was lower than with the expression of the mutant alone, but higher than with depletion of coronin 2A in the presence of wild-type Slingshot-1L.

These findings suggest a model where:

• Coronin-2A may regulate Slingshot-1L localization or activity

• It may enhance cofilin recruitment to specific actin structures

• Coronin-2A could protect certain actin structures while promoting turnover of others

This relationship explains how Coronin-2A coordinates with other actin-regulatory proteins to control cytoskeletal dynamics and cellular processes dependent on actin remodeling, particularly focal adhesion turnover during cell migration .

How does Coro2a contribute to endosomal fission and recycling pathways?

Coronin-2A plays a critical role in endosomal fission and recycling pathways through its interaction with the endosomal trafficking machinery. Research has identified specific mechanisms by which Coro2a influences these processes:

• EHD1 interaction: Coro2a contains an NPFDD sequence motif that preferentially binds to EHD1, a key component of the endosomal fission machinery . This interaction creates a molecular link between actin dynamics and membrane trafficking processes.

• Cargo-specific effects: Coro2a depletion causes differential effects on distinct cargo types:

  • Marked decrease in internalization of clathrin-dependent cargo (transferrin and EGF receptor)

  • Little impact on uptake of clathrin-independent cargo (MHC-1 and CD98 receptors)

  • Required for recycling of clathrin-independent cargo

• Endosomal morphology regulation: Coro2a depletion leads to enlarged endosomes, suggesting a direct role in the fission of endosomal vesicles . This enlargement likely results from impaired formation of transport vesicles from endosomal compartments.

• Actin-endosome coordination: Coro2a appears to coordinate actin-based endosomal processes with the EHD1 fission machinery, suggesting that actin dynamics play a role in endosomal vesicle formation and trafficking .

These findings establish Coro2a as a multifunctional protein that connects cytoskeletal regulation with membrane trafficking pathways. This coordination is essential for proper recycling of lipids and receptors to the plasma membrane, a process critical for maintaining cellular homeostasis .

What methodologies are most effective for studying Coro2a's role in regulating actin branching?

To effectively study Coronin-2A's role in regulating actin branching, researchers should employ multiple complementary methodologies:

• Visualization and quantification of actin structures:

  • Phalloidin staining to visualize F-actin distribution patterns

  • 3D filament reconstruction using specialized software (e.g., Imaris)

  • Automated quantification of actin-rich protrusions (size, frequency, distribution)

  • Super-resolution microscopy to examine detailed actin network architecture

• Molecular manipulation approaches:

  • siRNA or shRNA knockdown of Coro2a with appropriate controls

  • Rescue experiments with wild-type and mutant Coro2a constructs

  • CRISPR-Cas9 knockout for complete elimination of the protein

  • Expression of domain-specific mutants to dissect functional regions

• Dynamic analysis techniques:

  • Live-cell imaging with fluorescently tagged actin and Coro2a

  • FRAP (Fluorescence Recovery After Photobleaching) to measure actin turnover rates

  • Photoactivatable or photoconvertible actin to track specific populations

• Biochemical assays:

  • In vitro actin polymerization assays

  • Actin binding and bundling assays

  • Analysis of Arp2/3 complex activity in the presence/absence of Coro2a

The research has established that Coro2a depletion leads to dramatically expanded actin-rich protrusions, supporting its role in attenuating actin branching . These expanded protrusions increase both in size and frequency following Coro2a knockdown, similar to effects seen with other Coronin family proteins that disassemble Arp2/3-containing branched actin filaments.

Quantitative analysis using automated approaches is particularly valuable, as demonstrated by the application of 3D filament reconstruction tools that identify actin-rich protrusions as dendrite beginning points and calculate their frequency and mean diameter in an unbiased manner .

How might Coro2a function as a link between cytoskeletal dynamics and nuclear receptor signaling?

Coronin-2A may function as a molecular bridge between cytoskeletal dynamics and nuclear receptor signaling pathways. Research indicates that Coro2a serves as a key component of the nuclear receptor corepressor complex, which is essential for recruiting histone deacetylases to form repressive chromatin structures .

This dual functionality suggests several potential mechanisms:

• Cytoskeletal-nuclear signaling integration:

  • Coro2a may shuttle between cytoskeletal structures and nuclear complexes

  • Cytoskeletal changes could regulate Coro2a availability for nuclear functions

  • Coro2a might relay mechanical signals from the cytoskeleton to transcriptional machinery

• Coordinated regulation of gene expression and cell behavior:

  • Changes in cell adhesion or migration could be linked to specific transcriptional responses

  • Stress fibers, where Coro2a localizes, may function as mechanosensory structures

  • Nuclear receptor signaling might influence cytoskeletal organization through Coro2a

• Functional implications in development and disease:

  • This dual role could be crucial during developmental processes requiring coordination of cell movement and differentiation

  • Disruption of this link might contribute to diseases involving both cytoskeletal and transcriptional dysregulation

  • Cancer progression could be influenced by alterations in this cytoskeletal-nuclear connection

While direct experimental evidence for this link requires further investigation, Coro2a's presence in both actin-based structures and nuclear receptor complexes positions it as a potential integrator of physical and biochemical cellular processes . Future research using approaches like chromatin immunoprecipitation, nuclear/cytoplasmic fractionation, and targeted mutagenesis will help elucidate the functional significance of this dual localization.

What cell types and experimental models are most appropriate for studying Coro2a function?

Several cell types and experimental models have proven effective for studying different aspects of Coronin-2A function:

• Established cell lines:

  • HeLa cells: Effectively used for localization studies with FLAG-CORO2A

  • U251 Glioblastoma cells: Exhibit elaborate actin cytoskeletons ideal for studying actin organization

  • Various mouse cell lines: Appropriate for species-specific studies

• Primary cells:

  • Mouse embryonic fibroblasts: Useful for studying fundamental cytoskeletal functions

  • Immune cells: Potential models for investigating roles similar to other coronin family members

  • Neurons: Could reveal functions in specialized cell types with complex morphologies

• In vivo models:

  • Conditional knockout mice: Allow tissue-specific deletion of Coro2a

  • Transgenic models expressing tagged Coro2a: Enable in vivo tracking and localization

  • Developmental models: Could reveal roles in morphogenesis and tissue organization

• Disease models:

  • Cancer cell migration models: Given Coro2a's role in cell motility

  • Endocytosis/trafficking disorders: Based on its functions in endosomal processes

  • Metastasis models: To investigate potential roles in cancer progression

The choice of model system should be guided by the specific aspect of Coro2a function being investigated. For cytoskeletal studies, cells with well-developed actin networks like U251 are valuable , while trafficking studies benefit from cells with active endocytic pathways.

How does Coro2a function differ from other coronin family members?

Coronin-2A belongs to the type-II subfamily of coronin proteins and shows both similarities and distinct differences from other family members:

While both Coro2a and Type I coronins like Coronin-1B appear to attenuate actin branching, Coro2a has several unique functions:

• Its interaction with EHD1 through the NPFDD motif links it specifically to endosomal fission

• It serves as a component of the nuclear receptor corepressor complex

• It specifically regulates internal focal adhesion disassembly rather than peripheral adhesions

These distinctive properties suggest that despite structural similarities among coronin family members, they have evolved specialized functions in different cellular contexts.

What technological advances would enhance our understanding of Coro2a function?

Several technological advances would significantly enhance our understanding of Coronin-2A function:

• Advanced imaging techniques:

  • Super-resolution microscopy (STORM, PALM, STED) to visualize Coro2a interactions with actin and endosomes at nanoscale resolution

  • Lattice light-sheet microscopy for long-term 3D imaging of Coro2a dynamics in living cells

  • Correlative light and electron microscopy to connect fluorescence localization with ultrastructural context

• Protein interaction technologies:

  • BioID or APEX proximity labeling to identify the complete Coro2a interactome

  • Single-molecule tracking to measure binding kinetics in living cells

  • FRET-based biosensors to detect conformational changes or activation states

• Genome editing approaches:

  • CRISPR-Cas9 knock-in of fluorescent tags at endogenous loci

  • Domain-specific mutations at endogenous loci

  • Tissue-specific conditional knockout models

• Multi-omics integration:

  • Proteomics to identify Coro2a post-translational modifications

  • Transcriptomics to analyze effects on nuclear receptor target genes

  • Systems biology approaches to integrate multiple data types

• Structural biology techniques:

  • Cryo-EM structures of Coro2a-actin complexes

  • X-ray crystallography of the NPFDD motif interaction with EHD1

  • NMR studies of dynamic interactions and conformational changes

These technological advances would provide deeper insights into how Coro2a coordinates its diverse functions in actin regulation, focal adhesion turnover, endosomal trafficking, and potential nuclear roles.

What are the implications of Coro2a dysfunction in disease processes?

While the search results don't directly address disease associations, Coronin-2A's functions suggest several potential implications in disease processes:

• Cancer progression and metastasis:

  • Coro2a regulates cell motility, and its depletion reduces cell speed

  • Defects in focal adhesion turnover could affect cancer cell invasion

  • Altered expression might influence metastatic potential

• Trafficking-related disorders:

  • Coro2a's role in endosomal fission and recycling suggests potential involvement in:

    • Neurodegenerative diseases with trafficking defects

    • Disorders of receptor recycling or downregulation

    • Diseases involving defective protein sorting

• Developmental disorders:

  • If Coro2a coordinates cell migration with gene expression, its dysfunction could affect:

    • Tissue morphogenesis during development

    • Neural migration and connectivity

    • Organogenesis requiring coordinated cell movements

• Immune system dysfunction:

  • Given roles of other coronin family members in immune cells:

    • Coro2a might affect immune cell trafficking or function

    • Could influence inflammatory responses

    • Might play roles in immune surveillance

Research approaches to investigate these potential disease connections include:

• Analysis of Coro2a expression in disease tissues

• Correlation studies between Coro2a levels and disease progression

• Functional studies in disease-relevant cell types

• Development of animal models with tissue-specific Coro2a alterations

Understanding these disease implications could open new avenues for diagnostic and therapeutic strategies targeting Coro2a-related pathways.

What emerging research questions are most critical for advancing our understanding of Coro2a biology?

Several critical research questions would significantly advance our understanding of Coronin-2A biology:

• Mechanistic integration of diverse functions:

  • How does Coro2a coordinate its roles in actin regulation, focal adhesion turnover, and endosomal trafficking?

  • Are these functions regulated independently or interconnected through shared mechanisms?

  • Does Coro2a function as part of distinct protein complexes in different cellular contexts?

• Temporal and spatial regulation:

  • How is Coro2a activity regulated in time and space during dynamic cellular processes?

  • What post-translational modifications control its various functions?

  • How do cellular signals modulate its localization and interactions?

• Transcriptional regulation role:

  • What is the molecular basis of Coro2a's function in the nuclear receptor corepressor complex ?

  • Does Coro2a shuttle between cytoplasmic and nuclear compartments?

  • Which genes are regulated by Coro2a-containing complexes?

• Evolutionary conservation and specialization:

  • How have Coro2a functions evolved across species?

  • What unique properties distinguish it from other coronin family members?

  • Are there tissue-specific isoforms with specialized functions?

• Disease relevance:

  • How does Coro2a expression or function change in disease states?

  • Could targeting Coro2a-dependent pathways offer therapeutic opportunities?

  • Are there disease-associated mutations that affect Coro2a function?

Addressing these questions will require integrative approaches combining structural biology, advanced microscopy, proteomics, gene editing, and systems biology. The answers will provide a comprehensive understanding of how this multifunctional protein coordinates diverse cellular processes and may reveal new therapeutic targets for diseases involving cytoskeletal dysregulation or trafficking defects.

What are the key takeaways about Coro2a for researchers new to this field?

For researchers entering the field of Coronin-2A biology, several key concepts provide an essential foundation:

• Multifunctional protein: Coro2a is not simply an actin-binding protein but a multifunctional regulator involved in actin dynamics, focal adhesion turnover, endosomal trafficking, and potentially nuclear receptor signaling .

• Structural organization: As a type-II coronin, Coro2a contains an N-terminal β-propeller with WD40 repeats, a unique region, and a C-terminal coiled-coil domain that mediate its diverse interactions .

• Subcellular localization: Coro2a localizes predominantly to actin stress fibers (80% overlap with F-actin), with additional localization to focal adhesions and endosomal structures .

• Functional impacts: Depletion studies reveal that Coro2a:

  • Negatively regulates actin branching

  • Is essential for focal adhesion disassembly

  • Regulates endosomal fission and recycling

  • Controls cell motility

• Molecular interactions: Key interactions include binding to actin filaments, the endosomal protein EHD1 (via the NPFDD motif), and components of the nuclear receptor corepressor complex .

• Experimental approaches: Effective study of Coro2a involves:

  • Localization studies using tagged constructs

  • Knockdown/knockout approaches with appropriate controls

  • Rescue experiments to confirm specificity

  • Quantitative analysis of cytoskeletal and trafficking phenotypes

These fundamental aspects provide a starting point for designing experiments and interpreting results when investigating this complex protein in cellular systems.

What methodological recommendations can be made for researchers studying Coro2a?

Based on the published research, the following methodological recommendations would benefit researchers studying Coronin-2A:

• Protein detection and visualization:

  • Use validated antibodies like Coronin 2A Antibody (H-2) for western blotting and immunostaining

  • Employ both N- and C-terminally tagged fluorescent constructs to verify localization patterns

  • Include Phalloidin co-staining to confirm actin association

• Gene silencing approaches:

  • Implement multiple siRNA or shRNA sequences to control for off-target effects

  • Include rescue experiments with RNAi-resistant constructs

  • Validate knockdown by both protein and mRNA quantification

• Cell motility studies:

  • Combine scratch-wound assays with single-cell tracking for comprehensive analysis

  • Use kymography to analyze lamellipodial dynamics

  • Quantify specific parameters (speed, persistence, directionality)

• Cytoskeletal analysis:

  • Apply 3D reconstruction and automated quantification tools

  • Analyze specific parameters like protrusion size and frequency

  • Include cytochalasin D controls to confirm actin dependence

• Endosomal trafficking analysis:

  • Examine multiple cargo types (clathrin-dependent and independent)

  • Analyze both internalization and recycling processes

  • Quantify endosome morphology and size

• Controls and validation:

  • Include wild-type cells, mock-transfected cells, and non-targeting controls

  • Perform rescue experiments to confirm phenotype specificity

  • Use multiple cell types to verify consistency of findings

These methodological approaches will help ensure robust and reproducible results when investigating the diverse functions of Coronin-2A in cellular systems.