GCA Human

Grancalcin Human Recombinant
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Description

Introduction to Grancalcin (GCA Human)

Grancalcin (GCA), encoded by the GCA gene located on human chromosome 2q24.2 , is a calcium-binding protein belonging to the penta-EF-hand (PEF) subfamily of EF-hand proteins . It is predominantly expressed in neutrophils and macrophages, where it plays critical roles in calcium-dependent cellular processes, including granule-membrane fusion and immune response modulation .

Molecular Properties

Grancalcin exists as a single non-glycosylated polypeptide chain containing 237 amino acids (1–217.a.) and a molecular mass of 26.1 kDa . Recombinant human GCA is often expressed in E. coli with a 20-amino acid His-tag for purification .

Table 1: Molecular and Production Details of Grancalcin

ParameterValue/DescriptionSource
Amino Acid SequenceMGSSHHHHHHSSGLVPRGSH... (237 residues)
Molecular Mass26.1 kDa
SourceE. coli (recombinant)
Purity>95% (SDS-PAGE)
Formulation20 mM Tris-HCl (pH 8.0), 0.1 M NaCl, 20% glycerol

Functional Domains

Grancalcin contains five EF-hand motifs, which enable calcium binding and conformational changes . These structural features facilitate interactions with hydrophobic surfaces, such as neutrophil granules, and modulate adhesion to extracellular matrices like fibronectin .

Calcium-Dependent Localization

Grancalcin’s subcellular distribution is regulated by divalent cations:

  • No divalent cations: Cytoplasmic localization.

  • Magnesium alone: Granule-associated.

  • Calcium + Magnesium: Membrane- and granule-associated .

This dynamic localization suggests roles in granule secretion and cell adhesion during immune responses .

Interaction with L-Plastin

Grancalcin cooperates with L-plastin, an actin-bundling protein, to regulate neutrophil adhesion and migration . This interaction highlights its role in immune cell trafficking.

Comparative Analysis with PEF Proteins

Grancalcin shares structural and functional similarities with other PEF proteins:

ProteinKey FeaturesSource
GrancalcinNeutrophil adhesion, calcium-dependent fusion
CalpainCytoskeletal remodeling, protein degradation
SorcinCardiac calcium regulation, apoptosis
ALG-2Apoptosis regulation, endoplasmic reticulum

Gene and Epigenetic Insights

The GCA gene has been associated with 12 public variants in the Global Variome database, though disease links remain unclear . Epigenetic studies in vascular diseases like giant cell arteritis (GCA) highlight calcium-binding proteins’ roles in immune regulation , but direct links to Grancalcin are unexplored.

Table 2: Grancalcin’s Role in Neutrophil Function

ProcessMechanismEvidence
Granule SecretionCalcium-triggered membrane fusionLocalization studies
AdhesionInteraction with L-plastin and fibronectinFunctional assays

Product Specs

Introduction
Grancalcin (GCA) is a calcium-binding protein highly concentrated in human neutrophils. As a member of the penta EF-hand (PEF) protein family, which includes calpain, sorcin, peflin, and ALG-2, GCA undergoes crucial conformational changes upon calcium binding. This exposes hydrophobic amino acid residues, guiding the protein to hydrophobic surfaces. GCA interacts with L-plastin, known for its actin-bundling properties, suggesting a role for GCA in regulating neutrophil adhesion.
Description
Produced in E. coli, our GCA is a single, non-glycosylated polypeptide chain comprising 237 amino acids (1-217.a.) with a molecular weight of 26.1kDa. The protein is fused to a 20 amino acid His-tag at the N-terminus and purified using proprietary chromatographic techniques.
Physical Appearance
Sterile, clear, and colorless solution.
Formulation
The GCA protein solution (1mg/ml) is supplied in 20mM Tris-HCl buffer (pH 8.0), 0.1M NaCl, and 20% glycerol.
Purity
Purity exceeds 95% as determined by SDS-PAGE.
Stability
For short-term storage (up to 2-4 weeks), keep at 4°C. For extended storage, freeze at -20°C. Avoid repeated freeze-thaw cycles.
Synonyms
Grancalcin EF-hand calcium binding protein, GCL, Grancalcin penta-EF-hand protein.
Source
Escherichia Coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MAYPGYGGGF GNFSIQVPGM QMGQPVPETG PAILLDGYSG PAYSDTYSSA GDSVYTYFSA VAGQDGEVDA EELQRCLTQS GINGTYSPFS LETCRIMIAM LDRDHTGKMG FNAFKELWAA LNAWKENFMT VDQDGSGTVE HHELRQAIGL MGYRLSPQTL TTIVKRYSKN GRIFFDDYVA CCVKLRALTD FFRKRDHLQQ GSANFIYDDF LQGTMAI

Q&A

What is GCA and how should researchers operationalize it in study designs?

General Cognitive Abilities (GCA) refers to a set of mental capacities that enable individuals to reason, solve problems, comprehend complex ideas, learn quickly, and adapt to their environment. In research settings, GCA is typically operationalized through comprehensive assessment batteries rather than singular measures.

A methodologically sound approach to GCA assessment involves:

  • Employing standardized, validated test batteries that measure multiple cognitive domains

  • Ensuring tests have adequate psychometric properties (reliability ≥ 0.80, established construct validity)

  • Using age-appropriate normative data for score interpretation

  • Addressing cultural and linguistic biases through culturally-appropriate instruments

When designing studies, researchers should consider both specific and general factors of intelligence. Most contemporary research follows hierarchical models where specific cognitive abilities (e.g., processing speed, working memory) contribute to a higher-order general factor (g)1.

What are the essential methodological considerations for GCA assessment in diverse populations?

When conducting GCA research across diverse populations, methodological considerations must extend beyond basic psychometric properties. Researchers should implement:

  • Culture-fair assessment tools that minimize linguistic and cultural biases

  • Translation and back-translation procedures for cross-cultural research

  • Local normative data development when standard norms are inappropriate

  • Mixed-methods approaches that incorporate qualitative data for cultural context

  • Measurement invariance testing across groups to ensure valid comparisons

Testing conditions should be standardized but flexible enough to accommodate cultural differences. For example, in some cultures, speeded tests may introduce artificial performance deficits unrelated to actual cognitive abilities. Statistical approaches must account for demographic and socioeconomic variables that may influence cognitive performance differently across populations1 .

How can researchers effectively control for environmental variables in GCA studies?

Environmental variables present significant challenges to GCA research validity. Methodologically sound approaches include:

  • Documenting and controlling for socioeconomic status using validated measures

  • Assessing and accounting for educational quality and attainment

  • Measuring early life factors (nutrition, environmental exposures, trauma)

  • Implementing stratified sampling and matching procedures

  • Using statistical techniques such as propensity score matching or covariate adjustment

For experimental studies, researchers should standardize testing environments (time of day, noise levels, temperature) and counterbalance test order to control for fatigue effects. When field research is necessary, detailed documentation of environmental conditions allows for appropriate statistical control during analysis .

What ethical protocols must researchers follow when designing GCA studies?

GCA research requires rigorous ethical protocols beyond standard informed consent procedures:

  • Transparent communication about test purpose and interpretation limitations

  • Clear explanation of data storage, access, and confidentiality measures

  • Explicit description of potential risks (e.g., psychological distress from performance feedback)

  • Culturally sensitive procedures for obtaining informed consent

  • Plans for appropriate referral if clinically significant cognitive issues are detected

  • Procedures for reporting incidental findings that may impact participant health

Researchers must consider how cognitive assessment results might impact participants' self-perception and ensure debriefing procedures address these concerns. Special protections are required for vulnerable populations, including those with cognitive impairments that might affect capacity to consent .

What standard statistical approaches should researchers use when analyzing GCA data?

Statistical analysis of GCA data requires approaches that capture both measurement and theoretical complexity:

  • Structural equation modeling (SEM) for examining latent cognitive constructs

  • Hierarchical linear modeling for nested data (e.g., students within schools)

  • Factor analysis to identify underlying cognitive domains

  • Item response theory for test development and validation

  • Regression approaches with appropriate control variables

Researchers should report effect sizes and confidence intervals rather than relying solely on significance testing. When analyzing longitudinal GCA data, growth curve modeling and similar approaches that account for repeated measures are essential. For all analyses, researchers should address missing data appropriately through methods such as multiple imputation rather than simple deletion .

How should researchers address methodological challenges in longitudinal GCA studies?

Longitudinal GCA research presents unique methodological challenges requiring sophisticated approaches:

  • Implementing planned missing data designs to reduce participant burden while maximizing statistical power

  • Using modern psychometric techniques to address measurement invariance across time points

  • Employing appropriate statistical controls for practice effects

  • Accounting for cohort effects in accelerated longitudinal designs

  • Developing protocols to minimize attrition and techniques to address non-random dropout

When analyzing longitudinal GCA data, researchers should consider:

  • Growth mixture modeling to identify distinct cognitive trajectories

  • Multivariate latent growth curve models to examine relations between cognitive domains over time

  • Time-varying covariates to account for changing environmental influences

In developmental studies, age-appropriate assessment batteries must be carefully selected, with overlapping measures across age groups to enable continuous measurement across developmental periods1 .

How can researchers effectively integrate neuroimaging and genetic data with traditional GCA assessments?

Integration of neuroimaging and genetic data with traditional GCA measures requires multi-level analysis approaches:

  • Multimodal imaging protocols that capture both structural and functional correlates of cognition

  • Sophisticated statistical approaches like parallel ICA (Independent Component Analysis) to identify relationships between genetic variants and neuroimaging patterns

  • Mediation analyses to test whether neural mechanisms explain genetic influences on cognitive performance

  • Careful phenotyping with comprehensive cognitive batteries rather than single measures

Researchers should consider:

  • Sample size requirements for genetic and neuroimaging studies (typically larger than traditional behavioral studies)

  • Data harmonization procedures when combining datasets

  • Appropriate correction for multiple comparisons

  • Validation in independent samples

Integration of these complex data types requires interdisciplinary collaboration between cognitive scientists, neuroscientists, geneticists, and biostatisticians to ensure methodological rigor .

What methodological approaches help resolve data contradictions in GCA research?

When faced with contradictory findings in GCA research, methodologically sound approaches include:

  • Systematic analysis of measurement differences across studies

  • Evaluation of sample characteristics and potential moderating variables

  • Meta-analytic techniques to quantify heterogeneity and identify moderators

  • Multi-lab replication efforts with standardized protocols

  • Preregistration of study designs, hypotheses, and analysis plans

Researchers should systematically consider:

  • Measurement equivalence across studies

  • Population differences that might explain contradictory results

  • Methodological quality differences

  • Publication bias through techniques like funnel plots and p-curve analysis

When designing studies to resolve contradictions, researchers should implement designs that directly test competing hypotheses rather than simply replicating previous work with minor modifications1 .

How can researchers design valid experiments to test cognitive enhancement interventions?

Design of valid cognitive enhancement intervention studies requires:

  • Appropriate control conditions (active control vs. waitlist)

  • Adequate statistical power based on realistic effect size estimates

  • Comprehensive outcome measures assessing both near and far transfer

  • Follow-up assessments to determine maintenance of effects

  • Implementation of blinding procedures where possible

Researchers should address these methodological challenges:

  • Selection of appropriate control activities that match intervention characteristics without targeting the same cognitive mechanisms

  • Assessment of expectancy effects through self-report measures

  • Measurement of adherence and dosage

  • Thorough assessment of potential moderators (baseline cognitive ability, motivation, etc.)

Analysis approaches should include:

  • Intent-to-treat analyses alongside completer analyses

  • Examination of individual differences in response

  • Mediation analyses to identify mechanisms of change

  • Cost-effectiveness evaluation when appropriate1

What methodological approaches can address the replication crisis in GCA research?

To address replication challenges in GCA research, methodologically rigorous approaches include:

  • Pre-registration of study protocols, including specific hypotheses and analysis plans

  • Sample size planning based on realistic effect sizes rather than published estimates

  • Collaboration across multiple laboratories using identical protocols

  • Open data practices and code sharing

  • Transparent reporting of all analyses conducted (including unsuccessful analyses)

Specific methodological improvements include:

  • Reliability assessment for all measures with minimum thresholds

  • Establishing measurement invariance before making group comparisons

  • Reporting effect sizes with confidence intervals

  • Sequential testing approaches with alpha correction

  • Sensitivity analyses to determine the robustness of findings

Researchers should implement Registered Reports when appropriate, separating the evaluation of methods from results to reduce publication bias toward significant findings .

What methodological adaptations are required for GCA research in neurodevelopmental disorders?

GCA assessment in neurodevelopmental disorders requires methodological adaptations:

  • Selection of appropriate floor and ceiling levels to capture true variability

  • Administration modifications while maintaining standardization

  • Careful matching procedures for control groups (chronological vs. mental age)

  • Consideration of associated features (attention, motivation, sensory issues)

  • Integration of caregiver/teacher reports alongside direct assessment

When studying conditions like autism spectrum disorder, intellectual disability, or ADHD, researchers should:

  • Document medication status and consider washout procedures when appropriate

  • Assess domain-specific alongside general cognitive abilities

  • Consider adaptive functioning alongside cognitive performance

  • Select or develop measures validated for the specific population

  • Account for developmental trajectories in longitudinal designs

Statistical approaches should address potential floor and ceiling effects, non-normal distributions, and heterogeneity within diagnostic categories1 .

How should researchers approach GCA assessment in aging populations?

Methodologically sound approaches to studying GCA in aging populations include:

  • Comprehensive assessment of potential confounding health variables (cardiovascular health, medication use, etc.)

  • Use of measures validated for older adults with appropriate normative data

  • Careful attention to sensory limitations (vision, hearing) that might affect performance

  • Assessment of both crystallized and fluid abilities to capture differential age-related changes

  • Consideration of testing environment factors (fatigue, time of day effects)

When designing aging studies, researchers should:

  • Choose appropriate control groups (age-matched vs. young adults)

  • Account for cohort effects in cross-sectional designs

  • Conduct measurement invariance testing across age groups

  • Consider technology familiarity when using computerized assessment

  • Document retirement status, cognitive activities, and social engagement

Analysis approaches must address selective attrition in longitudinal studies, as healthier individuals are more likely to continue participation, potentially biasing results .

How can researchers effectively incorporate technological advances into GCA assessment?

Modern GCA research increasingly integrates technology through:

  • Computerized adaptive testing to optimize assessment efficiency

  • Virtual reality environments for ecologically valid assessment

  • Mobile cognitive assessment for experience sampling and ecological momentary assessment

  • Passive data collection through wearables and smartphone sensors

  • Machine learning approaches for pattern identification in complex datasets

Methodological considerations include:

  • Validation against traditional measures before widespread implementation

  • Evaluation of device effects (tablet vs. computer vs. smartphone)

  • Privacy and data security protocols

  • Consideration of digital literacy and access across populations

  • Standardization of hardware and software specifications

Researchers should pilot technological approaches with diverse samples to identify potential biases or accessibility issues before full implementation. Reporting should include detailed technical specifications to facilitate replication1 .

What methodological frameworks best support integration of cultural neuroscience in GCA research?

Cultural neuroscience approaches to GCA require methodological frameworks that:

  • Conceptualize culture beyond categorical variables (country, ethnicity)

  • Measure specific cultural values and practices that might influence cognitive development

  • Consider bidirectional relationships between neural development and cultural context

  • Implement mixed-methods approaches integrating qualitative and quantitative data

  • Apply appropriate cross-cultural neuroscience methods

Research designs should include:

  • Cultural adaptation of neuroscience paradigms

  • Validation of neuroimaging protocols across cultural contexts

  • Careful selection of comparison groups

  • Assessment of acculturation and biculturalism when relevant

  • Consideration of language effects on neural organization

Analysis approaches must test for measurement and structural invariance before making cross-cultural comparisons of brain-behavior relationships. Interdisciplinary collaboration with cultural anthropologists and cross-cultural psychologists strengthens methodological validity1 .

Product Science Overview

Structure and Function

Grancalcin contains four EF-hand domains, which are helix-loop-helix structural motifs capable of binding calcium ions. The protein undergoes significant conformational changes upon binding calcium, exposing hydrophobic amino acid residues that direct the protein to hydrophobic surfaces . This property is crucial for its role in cellular processes.

Localization and Role

The localization of grancalcin is dependent on the presence of divalent cations such as calcium and magnesium. In the absence of these cations, grancalcin is found in the cytosolic fraction. With magnesium alone, it partitions with the granule fraction, and in the presence of both magnesium and calcium, it associates with both the granule and membrane fractions . This suggests that grancalcin plays a role in granule-membrane fusion and degranulation, processes essential for the immune response.

Expression

Grancalcin is highly expressed in bone marrow and can be detected in neutrophils and macrophages . It is also expressed in other tissues such as the spleen, lung, and spinal ganglion .

Recombinant Grancalcin

Recombinant human grancalcin is produced using Escherichia coli expression systems. The recombinant protein typically includes a His-tag at the N-terminus to facilitate purification. The protein is purified using conventional chromatography techniques and is available in various formulations for research purposes .

Applications

Recombinant grancalcin is used in various research applications to study its role in calcium signaling, immune response, and cellular adhesion. It interacts with proteins such as L-plastin, which has actin-bundling activity, suggesting a role in the regulation of neutrophil adhesion .

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