CRYZL1 Human
Description
Protein Characteristics
CRYZL1 is a 40.8 kDa protein composed of 369 amino acids, produced recombinantly in E. coli as a non-glycosylated polypeptide. Key structural features include:
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N-terminal His-tag: Facilitates purification via nickel affinity chromatography .
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NAD(P)H binding site: Critical for its enzymatic activity as a quinone reductase .
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Sequence similarity: Shares homology with zeta-crystallin, a lens protein, but lacks the glycosylation patterns typical of crystallins .
| Parameter | Value/Description | Source |
|---|---|---|
| Molecular Weight | 40.8 kDa | |
| Amino Acids | 369 (1–349 + His-tag) | |
| Purity | >95% (SDS-PAGE) | |
| Storage Conditions | 4°C (short-term), -20°C (long-term) |
Enzymatic Activity
CRYZL1 exhibits NADPH-dependent quinone reductase activity, distinct from canonical quinone reductases. This activity suggests a role in detoxifying reactive quinones or modulating cellular redox states .
RNA Binding and Cellular Processes
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RNA interaction: Binds AU-rich elements in RNA, potentially regulating mRNA stability or translation .
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FERRY complex involvement: Part of the Five-subunit Endosomal Rab5 and RNA/ribosome intermediary (FERRY) complex, linked to endosomal trafficking and RNA processing .
Tissue Expression
CRYZL1 is expressed across multiple human tissues, with varying levels observed:
Mouse models show analogous expression patterns, including adipose tissue and hematopoietic systems .
Experimental Models
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Recombinant protein: Used in biochemical assays to study quinone reductase activity or RNA-binding kinetics .
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Single-cell profiling: Identifies cell-type-specific expression in tissues like the thymus, thyroid, and vascular systems .
Potential Therapeutic Targets
While no direct therapeutic applications are documented, its role in redox regulation and RNA processing suggests relevance in:
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Oxidative stress disorders: Neurodegeneration, cardiovascular disease.
Key Research Findings
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FERRY Complex Role: CRYZL1 interacts with Rab5 GTPase and ribosomal components, implicating it in endosomal trafficking and translation regulation .
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Alternative Splicing: Observed transcript variants, though full-length isoforms remain uncharacterized .
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pH Response Element Binding: Hypothesized role in pH-dependent cellular responses (e.g., stress adaptation) .
Product Specs
Q&A
What is the genomic organization of CRYZL1?
CRYZL1 is mapped to human chromosome 21q22.1 as confirmed by fluorescence in situ hybridization (FISH) . It appears to be a single-copy gene based on genomic Southern blot analysis with total human and yeast DNAs . The gene has alternatively spliced transcript variants, though their full-length nature has not been completely determined . To study genomic organization, researchers should employ a combination of:
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Whole genome sequencing with long-read technologies
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PCR-based exon mapping
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5' and 3' RACE (Rapid Amplification of cDNA Ends) to identify all transcript variants
What is the tissue expression profile of CRYZL1?
CRYZL1 mRNA is expressed in multiple human tissues including heart, brain, skeletal muscle, kidney, pancreas, liver, and lungs, though at varying levels across these tissues . To comprehensively analyze tissue-specific expression patterns, researchers should:
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Use quantitative RT-PCR with tissue-specific RNA samples
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Perform Western blot analysis with validated antibodies
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Employ immunohistochemistry for spatial localization within tissues
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Consider single-cell RNA sequencing for cell-type specific expression patterns
What is the enzymatic function of CRYZL1?
CRYZL1 encodes a protein with sequence similarity to zeta crystallin, also known as quinone oxidoreductase . The protein contains an NAD(P)H binding site, suggesting it may catalyze redox reactions involving quinone substrates . To characterize enzymatic function, researchers should:
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Express and purify recombinant CRYZL1 protein
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Perform enzyme kinetics assays with various quinone substrates
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Measure NAD(P)H consumption using spectrophotometric methods
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Conduct site-directed mutagenesis of the NAD(P)H binding domain to confirm functional importance
How does CRYZL1 interact with other proteins and pathways?
The search results indicate CRYZL1 mRNA can interact with G3BP1 protein , suggesting potential post-transcriptional regulation during stress responses. Additionally, the RORA protein (retinoic acid receptor-related orphan receptor A) has been reported to bind to the CRYZL1 gene . To map protein-protein interactions:
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Perform co-immunoprecipitation followed by mass spectrometry
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Use yeast two-hybrid screening
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Employ proximity labeling methods (BioID, APEX)
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Validate interactions with co-localization studies
How do environmental chemicals affect CRYZL1 expression?
CRYZL1 expression is modulated by numerous environmental compounds, as evidenced by extensive chemical interaction data. The following table summarizes key compounds affecting CRYZL1:
To investigate chemical effects on CRYZL1, researchers should:
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Design dose-response and time-course experiments
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Use reporter gene assays with CRYZL1 promoter constructs
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Employ ChIP-seq to identify transcription factor binding changes
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Analyze methylation patterns with bisulfite sequencing
What methodologies best detect CRYZL1 epigenetic modifications?
Several compounds affect CRYZL1 methylation, including aflatoxin B1 (increases methylation) and titanium dioxide (decreases methylation) . For studying epigenetic regulation:
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Use bisulfite conversion followed by sequencing
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Employ methylation-specific PCR
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Conduct chromatin immunoprecipitation (ChIP) with antibodies against histone modifications
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Perform ATAC-seq to assess chromatin accessibility at the CRYZL1 locus
What is the potential role of CRYZL1 in Down syndrome?
Given CRYZL1's location on chromosome 21q22.1 , it may be overexpressed in Down syndrome due to trisomy 21. To investigate its contribution to Down syndrome pathophysiology:
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Compare CRYZL1 expression levels between Down syndrome and control samples
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Create cellular models with controlled CRYZL1 overexpression
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Analyze downstream effects on redox homeostasis and cellular metabolism
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Investigate protein interaction networks in the context of other chromosome 21 genes
How might CRYZL1 contribute to oxidative stress-related disorders?
As a potential quinone oxidoreductase, CRYZL1 likely influences cellular redox balance. To study its role in oxidative stress conditions:
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Use CRISPR/Cas9 to create CRYZL1 knockout or overexpression models
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Measure reactive oxygen species (ROS) levels using fluorescent probes
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Assess cell viability under oxidative challenge
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Analyze metabolic changes using targeted metabolomics
How can contradictory findings about CRYZL1 expression be reconciled?
The search results indicate contradictory findings regarding CRYZL1 expression in response to certain compounds (e.g., valproic acid both increases and decreases expression) . To resolve such contradictions:
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Design comprehensive time-course experiments to detect temporal expression dynamics
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Conduct dose-response studies to identify threshold effects
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Perform cell type-specific analyses to account for tissue-specific responses
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Investigate context-dependent regulatory mechanisms through multi-omics approaches
What cutting-edge technologies would advance CRYZL1 functional studies?
To propel CRYZL1 research forward, researchers should consider:
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Cryo-EM for high-resolution structural analysis of CRYZL1 protein
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Proteome-wide thermal shift assays to identify binding partners
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CRISPR screens to identify synthetic lethal interactions
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Single-cell multi-omics to correlate CRYZL1 expression with cellular phenotypes
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Spatial transcriptomics to map expression patterns within complex tissues
What are the critical controls for CRYZL1 expression studies?
When studying CRYZL1 expression, researchers should implement:
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Positive and negative control tissues based on known expression patterns in heart, brain, muscle, kidney, pancreas, liver, and lung
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Multiple reference genes for qPCR normalization
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Antibody validation using CRYZL1 knockout or knockdown samples
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Inclusion of alternatively spliced variants in primer design
How should researchers address potential functional redundancy?
Given CRYZL1's similarity to other quinone oxidoreductases, functional redundancy is a significant consideration. To address this:
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Create combinatorial knockouts of CRYZL1 and related enzymes
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Conduct comparative substrate specificity assays
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Perform rescue experiments with various family members
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Use systems biology approaches to map metabolic network adaptations
Product Science Overview
The CRYZL1 gene is located on chromosome 13 in humans . The gene undergoes alternative splicing, resulting in multiple transcript variants, although the full-length nature of these variants has not been completely determined . The protein encoded by this gene is involved in various cellular processes, including the reduction of quinones to hydroquinones, which helps in detoxifying reactive quinones and preventing oxidative stress .
Quinone oxidoreductase-like protein 1 functions as a two-electron reductase, catalyzing the reduction of quinones to hydroquinones . This reaction is essential for detoxifying quinones, which can generate reactive oxygen species (ROS) through redox cycling mechanisms . By reducing quinones to hydroquinones, CRYZL1 helps in preventing the formation of semiquinone free radicals and other harmful oxidative species .
The physiological role of CRYZL1 extends beyond its enzymatic activity. It is involved in maintaining cellular redox balance and protecting cells from oxidative damage . The enzyme’s activity is crucial in tissues exposed to high levels of oxidative stress, such as the liver and lungs . Additionally, CRYZL1 plays a role in the metabolism of vitamin K, which is essential for blood clotting and bone health .
Mutations or alterations in the CRYZL1 gene can have significant clinical implications. For instance, certain polymorphic forms of the protein may be associated with an increased risk of developing various diseases, including cancer . The enzyme’s role in detoxifying reactive quinones and maintaining cellular redox balance makes it a potential target for therapeutic interventions in diseases characterized by oxidative stress and redox imbalance .
The human recombinant form of Quinone Oxidoreductase-like Protein 1 is produced using recombinant DNA technology. This involves inserting the CRYZL1 gene into an expression vector, which is then introduced into a host cell (such as E. coli or yeast) to produce the protein . The recombinant protein is purified and used in various research applications, including studies on enzyme function, drug development, and disease mechanisms .
