Podcast
Questions and Answers
What is the main goal of protein engineering?
What is the main goal of protein engineering?
Which of the following is a key advantage of directed evolution?
Which of the following is a key advantage of directed evolution?
Which of the following best describes rational design?
Which of the following best describes rational design?
Which amino acid contains a thiol group useful for bioconjugation?
Which amino acid contains a thiol group useful for bioconjugation?
Signup and view all the answers
Which of these is not a post-translational modification?
Which of these is not a post-translational modification?
Signup and view all the answers
What is the primary limitation of Solid-Phase Peptide Synthesis (SPPS)?
What is the primary limitation of Solid-Phase Peptide Synthesis (SPPS)?
Signup and view all the answers
Which of the following describes a key feature of Cell-Free Protein Synthesis (CFPS)?
Which of the following describes a key feature of Cell-Free Protein Synthesis (CFPS)?
Signup and view all the answers
What is a primary advantage of recombinant DNA technology in protein synthesis?
What is a primary advantage of recombinant DNA technology in protein synthesis?
Signup and view all the answers
What is the most common glycosylation type in antibodies?
What is the most common glycosylation type in antibodies?
Signup and view all the answers
Which PTM stabilizes protein structure via covalent bonding?
Which PTM stabilizes protein structure via covalent bonding?
Signup and view all the answers
Which PTM adds a negatively charged group to proteins?
Which PTM adds a negatively charged group to proteins?
Signup and view all the answers
Which of the following PTMs typically regulates transcription?
Which of the following PTMs typically regulates transcription?
Signup and view all the answers
What is the target functional group of NHS esters in bioconjugation?
What is the target functional group of NHS esters in bioconjugation?
Signup and view all the answers
Which reagent is commonly used for thiol-specific cross-linking?
Which reagent is commonly used for thiol-specific cross-linking?
Signup and view all the answers
What is the purpose of bifunctional cross-linkers like NHS-PEG-Maleimide?
What is the purpose of bifunctional cross-linkers like NHS-PEG-Maleimide?
Signup and view all the answers
Which type of reaction is typically used for hydroxyl groups in tyrosine?
Which type of reaction is typically used for hydroxyl groups in tyrosine?
Signup and view all the answers
Which functional group in histidine enables metal coordination?
Which functional group in histidine enables metal coordination?
Signup and view all the answers
What is the principle of Top-Down Mass Spectrometry?
What is the principle of Top-Down Mass Spectrometry?
Signup and view all the answers
How does SILAC enable quantification of proteins?
How does SILAC enable quantification of proteins?
Signup and view all the answers
Which tool is most commonly used for structural prediction in protein engineering?
Which tool is most commonly used for structural prediction in protein engineering?
Signup and view all the answers
Which of the following describes a LOCKR protein system?
Which of the following describes a LOCKR protein system?
Signup and view all the answers
What is the role of fluorescent protein bioconjugation to antibodies?
What is the role of fluorescent protein bioconjugation to antibodies?
Signup and view all the answers
Why is click chemistry considered bioorthogonal?
Why is click chemistry considered bioorthogonal?
Signup and view all the answers
Which of the following is a unique feature of cell-free protein synthesis?
Which of the following is a unique feature of cell-free protein synthesis?
Signup and view all the answers
Which amino acid is most commonly glycosylated in antibodies?
Which amino acid is most commonly glycosylated in antibodies?
Signup and view all the answers
What is a common use of periodate oxidation?
What is a common use of periodate oxidation?
Signup and view all the answers
Which PTM is commonly used to target proteins for degradation?
Which PTM is commonly used to target proteins for degradation?
Signup and view all the answers
Which cross-linking reagent is a zero-length cross-linker?
Which cross-linking reagent is a zero-length cross-linker?
Signup and view all the answers
What is the effect of phosphorylation on protein charge?
What is the effect of phosphorylation on protein charge?
Signup and view all the answers
What is a key limitation of directed evolution?
What is a key limitation of directed evolution?
Signup and view all the answers
Study Notes
Protein Engineering Exam MCQs - General Concepts
- Protein engineering goal: Create proteins with desired functions
- Directed evolution advantage: Does not require prior structural knowledge
- Rational design key: Detailed structural and functional knowledge of the protein is needed
- Bioconjugation amino acid: Cysteine (contains a thiol group)
- Post-translational modification (not): Recombinant DNA insertion
- Rational design: Mimics natural selection in vitro, needs detailed structural & functional knowledge of the protein
- Directed evolution: Requires no prior structural knowledge; doesn't necessarily yield a perfect protein in one cycle; utilizes non-natural amino acids, but screening is still required
Protein Synthesis
- Solid-Phase Peptide Synthesis (SPPS) limitation: Difficulty synthesizing large proteins
- Cell-Free Protein Synthesis (CFPS) key feature: Enables rapid incorporation of non-natural amino acids
- Recombinant DNA advantage: Enables large-scale protein production in host cells.
- Recombinant DNA primary advantage: Allows large-scale protein production in host cells - avoids problems of using live cells
- Primary limitations of SPPS (Solid Phase Peptide Synthesis): Inability to incorporate non-natural amino acids, difficulty in synthesizing large proteins, requirement of living cells for synthesis, and lack of precision in amino acid coupling
Post-Translational Modifications (PTMs)
- Common Antibody Glycosylation type: N-linked glycosylation
- Stabilizes Protein Structure: Disulfide bond formation (via covalent bonding)
- Negatively charged group addition: Phosphorylation
- Transcription regulation PTM: Ubiquitination
Bioconjugation and Cross-Linking
- NHS esters target functional group: Amine (-NH2)
- Thiol-specific cross-linking agent: Maleimide
- Bifunctional cross-linker purpose: To link two different types of functional groups
- Hydroxyl group reaction: Periodate oxidation
- Metal coordination: Imidazole group (in histidine)
Techniques in Protein Engineering
- Top-Down Mass Spectrometry principle: Fragmentation/analysis of intact proteins
- SILAC protein quantification: Uses isotopically labeled amino acids for synthesis.
- Structural prediction tool: AlphaFold
- LOCKR protein system: A method to create isotopically labeled proteins
- Fluorescent protein bioconjugation use with Antibodies: Enables detection and visualization
- Click chemistry: Bioorthogonal, meaning it works without interfering with native biomolecules in living systems
- Cell-free protein synthesis advantage: Eliminates the need for transcription and translation machinery
- Commonly glycosylated amino acid in antibodies: Asparagine
Advanced Concepts/Questions
- Zero-length cross-linker: EDC
- Phosphorylation effect on protein charge: Introduces a negative charge
- Directed evolution limitation: Requires extensive structural knowledge, screening is resource-intensive
- Common use for periodate oxidation: Introducing thiol groups into proteins.
- PTM commonly used for protein degradation: Ubiquitination.
Studying That Suits You
Use AI to generate personalized quizzes and flashcards to suit your learning preferences.
Related Documents
Description
This quiz focuses on the fundamental concepts of protein engineering, including the goals of creating proteins with desired functions and the methodologies like directed evolution and rational design. Test your understanding of protein synthesis techniques such as Solid-Phase and Cell-Free Protein Synthesis. Perfect for those studying biochemical and molecular biology.
