GPT2 Human

Glutamic-Pyruvate Transaminase 2 Human Recombinant
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Description

Gene and Protein Structure

  • Chromosomal Location: 16q11.2 .

  • Protein:

    • 546 amino acids (1–523 residues) with a molecular mass of 60.3 kDa .

    • Contains a catalytic aspartate aminotransferase (AAT) domain critical for enzymatic activity .

    • Post-translational modifications include a 23-amino acid His-tag in recombinant forms .

Functional Domains

  • Catalytic activity depends on pyridoxal phosphate (PLP) binding .

  • Key mutations (e.g., p.Arg404*, p.Pro272Leu) impair enzyme activity, substrate binding, or protein stability .

Biochemical Properties

Recombinant GPT2 Human (produced in E. coli) is commercially available for research. Key specifications include:

PropertyDetails
SourceE. coli expression system
Purity>90% (SDS-PAGE)
Formulation20 mM Tris-HCl (pH 7.5), 30% glycerol, 2 mM DTT, 0.2 M NaCl
StabilityStore at -20°C; avoid freeze-thaw cycles

Functional Role in Human Physiology

GPT2 Human facilitates:

  • Amino Acid Metabolism: Converts glutamate to α-ketoglutarate, linking nitrogen metabolism to the TCA cycle .

  • Anaplerosis: Replenishes TCA intermediates critical for mitochondrial energy production .

  • Neuroprotection: Regulates antioxidants and neuroprotective metabolites in the brain .

In Vitro Studies

  • Hypoxia Response: Hypoxia-inducible factor 2 (HIF-2) upregulates GPT2 in glioblastoma (GBM), enhancing cell migration via α-ketoglutarate modulation .

  • Enzyme Activity: Missense/nonsense mutations (e.g., Q80E, C259R) reduce catalytic activity by >90% .

In Vivo Models

  • Mouse Models: Gpt2-null mice exhibit:

    • Postnatal microcephaly and motor deficits .

    • Metabolic disruptions: low alanine, elevated essential amino acids (e.g., phenylalanine) .

  • Cancer: GPT2 ablation in triple-negative breast cancer inhibits tumor growth by impairing glutaminolysis and inducing autophagy .

Genetic Disorders

  • GPT2-Syndrome: Autosomal recessive disorder characterized by intellectual disability, microcephaly, and spastic paraplegia .

    • Mutations: 12 pathogenic variants reported, primarily in the AAT domain .

    • Diagnostic Testing: Clinical sequencing identifies homozygous or compound heterozygous mutations .

Therapeutic Targets

  • Metabolic interventions (e.g., α-ketoglutarate supplementation) may mitigate symptoms .

  • HIF-2 inhibitors show potential in GPT2-driven cancers .

Comparative Analysis of GPT2 Mutations

MutationTypeFunctional ImpactPhenotype
p.Arg404*NonsensePremature truncation; loss of catalytic domainSevere encephalopathy, motor deficits
p.Pro272LeuMissenseDisrupted substrate bindingDevelopmental delay, microcephaly
V478Rfs*73FrameshiftAltered protein foldingProgressive spastic paraplegia

Future Directions

  • Investigate GPT2’s role in neurodegenerative diseases (e.g., ALS, Parkinson’s) .

  • Develop isoform-specific inhibitors for cancer therapy .

Product Specs

Introduction
Alanine aminotransferase 2 (GPT2) is an enzyme that plays a crucial role in the metabolism of glucose and amino acids. It catalyzes the reversible conversion of alanine and 2-oxoglutarate to pyruvate and glutamate. GPT2 is primarily found in muscle, fat, and kidney tissues. Several different forms of GPT2, known as isoforms, are produced from multiple transcript variants.
Description
Recombinant human GPT2 is a 60.3 kDa protein containing 546 amino acids (specifically, amino acids 1-523). Produced in E. coli, this single polypeptide chain is fused to a 23 amino acid His-tag at its N-terminus and purified using proprietary chromatographic techniques.
Physical Appearance
Clear, colorless, and sterile-filtered solution.
Formulation
The GPT2 solution is provided at a concentration of 0.5 mg/ml and contains the following components: 20mM Tris-HCl buffer (pH 7.5), 30% glycerol, 0.2M NaCl, and 2mM DTT.
Stability
For short-term storage (2-4 weeks), the GPT2 solution should be kept at 4°C. For long-term storage, it is recommended to store the solution frozen at -20°C. Adding a carrier protein such as 0.1% HSA or BSA is advisable for extended storage durations. Repeated freezing and thawing of the solution should be avoided.
Purity
The purity of the GPT2 protein is greater than 90%, as determined by SDS-PAGE analysis.
Synonyms
ALT2, AAT2, Alanine aminotransferase 2, Glutamate pyruvate transaminase 2, Glutamic--alanine transaminase 2, Glutamic--pyruvic transaminase 2.
Source
E.coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MGSMQRAAAL VRRGCGPRTP SSWGRSQSSA AAEASAVLKV RPERSRRERI LTLESMNPQV KAVEYAVRGP IVLKAGEIEL ELQRGIKKPF TEVIRANIGD AQAMGQQPIT FLRQVMALCT YPNLLDSPSF PEDAKKRARR ILQACGGNSL GSYSASQGVN CIREDVAAYI TRRDGGVPAD PDNIYLTTGA SDGISTILKI LVSGGGKSRT GVMIPIPQYP LYSAVISELD AIQVNYYLDE ENCWALNVNE LRRAVQEAKD HCDPKVLCII NPGNPTGQVQ SRKCIEDVIH FAWEEKLFLL ADEVYQDNVY SPDCRFHSFK KVLYEMGPEY SSNVELASFH STSKGYMGEC GYRGGYMEVI NLHPEIKGQL VKLLSVRLCP PVSGQAAMDI VVNPPVAGEE SFEQFSREKE SVLGNLAKKA KLTEDLFNQV PGIHCNPLQG AMYAFPRIFI PAKAVEAAQA HQMAPDMFYC MKLLEETGIC VVPGSGFGQR EGTYHFRMTI LPPVEKLKTV LQKVKDFHIN FLEKYA.

Q&A

What distinguishes GPT-2 generated content from human-written material in experimental studies?

Research comparing GPT-2 generated content with human-written material has revealed several interesting distinctions:

CharacteristicGPT-2 Generated ContentHuman-Written Content
Plausibility87.59% contain globular domains88.40% contain globular domains
Secondary Structure48.64% alpha-helical, 39.70% beta-sheet45.19% alpha-helical, 41.87% beta-sheet
Preference RatingOften preferred when conveying similar messagesSometimes less preferred in direct comparisons

Interestingly, in a study examining pairs of similar claims, human annotators preferred the GPT-2 version over the human-authored one in 13 out of 20 cases, with 4 cases rated equally and only 4 cases where the human version was preferred . This suggests that GPT-2 can sometimes produce more appealing phrasings than humans, even when conveying the same information.

What experimental designs are most effective for evaluating human perception of GPT-2 outputs?

Several experimental designs have proven effective for evaluating human perception of GPT-2 generated content:

  • Comparative Evaluation: Presenting human evaluators with pairs of outputs (GPT-2 vs. human) and asking which is preferred

  • Plausibility Assessment: Determining whether generated content could plausibly have been created by a human

  • Cascaded Evaluation: First assessing plausibility, then only evaluating stance or other attributes for plausible outputs

  • Multi-dimensional Rating: Evaluating outputs across multiple quality dimensions (coherence, factuality, relevance)

  • Automatic + Manual Hybrid: Combining automatic metrics (e.g., stance detection scores) with human judgments

Research has shown that cascaded evaluation approaches can be particularly efficient, as they filter out low-quality outputs early in the process . This methodological approach ensures that research resources are focused on the most promising outputs.

How can researchers implement reinforcement learning from human feedback (RLHF) with GPT-2?

Implementing RLHF with GPT-2 requires a systematic methodology:

  • Preparation Phase:

    • Fine-tune GPT-2 on your target domain

    • Create a dataset of relevant prompts

    • Establish annotation guidelines

  • Data Collection:

    • Generate multiple completions per prompt

    • Collect human preferences between samples

    • Ensure diverse preference data

  • Reward Model Training:

    • Train a model to predict human preferences

    • Validate against held-out preference data

  • Policy Optimization:

    • Use PPO to optimize the language model

    • Include KL-divergence penalty to prevent divergence

    • Continuously evaluate and iterate

OpenAI's research demonstrated that this methodology successfully aligns GPT-2 with human preferences, though interestingly, "those preferences did not always match our own" . This highlights the importance of carefully designing preference collection protocols that capture the desired values and objectives.

How do different fine-tuning datasets affect GPT-2's ability to generate human-like content?

Research has examined the impact of different datasets on GPT-2's ability to generate human-like content:

Interestingly, research has found that "manually labeled datasets used to fine-tune GPT-2 are not essential, and that relying on the output of a Claim Retrieval engine for this fine-tuning, may suffice" . This suggests that researchers might be able to leverage existing tools and datasets to create effective fine-tuning data with reduced manual labeling costs.

What are the key methodological considerations for collecting high-quality human preference data?

Collecting high-quality human preference data for GPT-2 research involves several methodological considerations:

  • Task Design:

    • Clear instructions and evaluation criteria

    • Appropriate cognitive load and task complexity

    • Balanced exposure to different types of content

  • Annotator Selection and Training:

    • Diverse annotator demographics

    • Comprehensive training with examples

    • Quality control mechanisms

  • Data Collection Format:

    • Pairwise comparisons vs. Likert scales

    • Number of options presented simultaneously

    • Context provided to annotators

  • Quality Assurance:

    • Inter-annotator agreement monitoring

    • Attention checks and calibration examples

    • Iterative refinement of instructions

  • Bias Mitigation:

    • Randomization of presentation order

    • Balanced content selection

    • Monitoring for systematic biases

Research has shown that the specific choice of data collection methodology can significantly impact the resulting model behavior . For example, asking labelers to compare four samples (rather than pairs) may provide more efficient preference signals, while ensuring accuracy checks in summarization tasks may lead to different preference patterns.

How can researchers balance dataset size requirements with human labeling costs?

Researchers can employ several strategies to optimize the trade-off between dataset size and labeling costs:

  • Task-Appropriate Scaling: Simpler tasks (like stylistic continuation) may require only ~5,000 human samples, while complex tasks (like summarization) might need ~60,000 labels .

  • Active Learning: Selecting the most informative samples for human annotation rather than random sampling.

  • Semi-Supervised Approaches: Using small amounts of human-labeled data supplemented with model-generated data.

  • Cascade Filtering: Using automated methods (like claim detection tools) to filter generated outputs before human evaluation .

  • Transfer Learning: Fine-tuning models on related tasks first to reduce target task data requirements.

The optimal strategy depends on task complexity, available resources, and required performance levels. Research indicates that post-generation ranking steps can significantly enhance quality, suggesting that efficient filtering of generated outputs may be more cost-effective than collecting more training data .

What statistical methods are most appropriate for analyzing human preference data?

Several statistical approaches are particularly suited for analyzing human preference data in GPT-2 research:

  • Preference Learning Models:

    • Bradley-Terry model for pairwise comparisons

    • Plackett-Luce model for ranking data

    • These models estimate underlying quality scores from preference data

  • Agreement Metrics:

    • Cohen's Kappa for categorical judgments

    • Krippendorff's Alpha for multiple annotators

    • These measure reliability and consistency in human judgments

  • Comparative Analysis Techniques:

    • Significance testing between different model versions

    • Confidence intervals for preference proportions

    • These help quantify the reliability of observed differences

  • Mixed-Effects Models:

    • Account for annotator-specific and item-specific random effects

    • Control for confounding variables

    • Provide more nuanced analysis of preference patterns

Product Science Overview

Gene and Protein Structure

The GPT2 gene is located on chromosome 16 at the position 16q11.2 . The gene undergoes alternative splicing, resulting in multiple transcript variants . The protein encoded by GPT2 is a pyridoxal enzyme, which means it requires pyridoxal phosphate (a form of vitamin B6) as a cofactor for its enzymatic activity .

Function and Biological Role

GPT2 is primarily involved in the catabolism of L-alanine and the biosynthesis of cellular amino acids . It is expressed in various tissues, including skeletal muscle, liver, kidney, and pancreas . The enzyme’s activity is crucial for maintaining the balance of amino acids and for the production of glucose from non-carbohydrate sources, a process known as gluconeogenesis .

Under metabolic stress conditions, the expression of GPT2 is upregulated by activating transcription factor 4 (ATF4) in hepatocyte cell lines . This regulation ensures that the enzyme is available to meet the increased metabolic demands during stress.

Clinical Significance

Mutations in the GPT2 gene have been associated with developmental encephalopathy, a condition characterized by severe developmental delays and neurological abnormalities . Additionally, GPT2 is linked to other metabolic disorders, such as Neurodevelopmental Disorder with Spastic Paraplegia and Microcephaly and Glycogen Storage Disease III .

Research and Applications

Recombinant forms of GPT2 are used in research to study its structure, function, and role in various metabolic pathways. These studies help in understanding the enzyme’s involvement in metabolic diseases and in developing potential therapeutic interventions.

In summary, Glutamic-Pyruvate Transaminase 2 (Human Recombinant) is a vital enzyme with significant roles in amino acid metabolism and gluconeogenesis. Its regulation and function are crucial for maintaining metabolic homeostasis, and mutations in the GPT2 gene can lead to severe metabolic disorders.

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