VISHWA BHARATI PUBLIC SCHOOL, NOIDA
HOLIDAY HOMEWORK
CLASS XII
Topic : Protein Structure & Engineering
Skill : Knowledge, Reasoning & Application
Level : Average & above average
Type : Assignment
General instructions:
1. Attempt all one mark questions in the worksheet itself.
2. Time yourself as per the expected time given against each type of question.
3. Supplement your answer with neat, well-labelled diagrams where ever possible.
4. Use of calculators is not allowed, however you can use log books if required.
5. Paste this worksheet in your register.
I Very Short Answer Questions (1 mark each) Expected time/question: 1- 2 minutes
1. Give one example each of Biomolecules involved in the process of:
(i) Muscular expansion & contraction ………………………………………………………………
(ii) Body structure & protection ………………………………………………………………
2. ………………. are linear polymers of amino acids linked together by ……………………… formed between the ……………………………….. and …………...…………...…… of two adjacent amino acids.
3. Protein concentration is preferably monitored at ……………………………………………… wavelength.
4. ……………………….. and ……………………….. are two reagents commonly used for protein structure elucidation.
5. ………………………………………… was the first protein to be sequenced.
6. To date only …………………………. proteins have been identified.
7. Glutathione helps in ………………………………………………………………………………………….
8. Nitrogen part of BCAA is used for….………………………………………………………………………..
9. Carbon part of BCAA is used for…………………………………………………………………………….
10. ……………………………...……… and ……………………………….……….. are muscular proteins.
11. The chemical component that gives proper shape to the living cell is ……...……………………………….
12. What happens if:
(i) A protein misfolds/undergoes conformational changes ……...... ……………………………………
(ii) Lack of beta chain in hemoglobin ……..………………………………………..
(iii) Presence of mutated beta resulting in erythrocytes ………………………………………………
(iv) Absence of adenosine deaminase ………………………………………………
13. ………………………………… interactions determine the shape of the protein.
14. Active sites exposed on protein surface by…………………………………………………………………...
15. ………………….….… and …………..……...….. structures represent the most thermodynamically stable conformation of protein is in solution and result from ……………………….………. interactions between …………………… side chains within the molecule and with the ……………… molecules surrounding it.
16. …………………………………………………….. is a commonly used dye to stain proteins.
17. Define the following terms:
(i) Epitope (ii) Parental preparation
(iii) Receptor proteins (iv) Probiotics
18. Give the contributions of the following scientists:
(i) Pehr Edman ………………………………………………………………………………………
(ii) Fredrick Sanger ………………………………………………………………………………………
(iii) Pauling, Ramachandran, Perutz & Kendrew ………………………………………………………
(iv) Linus Pauling ………………………………………………………………………………………
(v) V.M.Ingram ………………………………………………………………………………………
(vi) O’Farrel ………………………………………………………………………………………
(vii) Baricelli ………………………………………………………………………………………
19. Give an example of each of the following:
(i) A protein having primary structure ………………………………………………………………
(ii) A protein having secondary structure ………………………………………………………………
(iii) A protein having tertiary structure ………………………………………………………………
(iv) A protein having quaternary structure ………………………………………………………………
(v) Amino acid having hydrophobic group ………………………………………………………………
(vi) Amino acid having polar group ………………………………………………………………
(vii) Amino acid having charged group ………………………………………………………………
(viii) Intracellular protein ………………………………………………………………
(ix) Extracellular protein ………………………………………………………………
20. When we refer to the structure of protein it involves the …………………………………………… and the …………………………………………..
21. Give the mechanism of action of the following:
(i) Chymotrypsin …...………………………………………………………………………………….
(ii) Trypsin ………………………………………………………………..…………….……….
(iii) Subtilisin ………………………………………………………………………………………
(iv) Hexokinase ………………………………………………………………………………………
22. Isolation of a protein from a microbe, plant or animal cell requires various separation techniques collectively known as ………………………………………….
23. Name any one technique used for the obtaining the following:
(i) Protein structural information ………………………………………
(ii) Amino acid sequences ………………………………………
(iii) Analyzing high mol wt compound ………..……………………………..
(iv) Gaseous ionized molecules from solution ………………………………………
(v) Separation of cellular debris from soluble protein ………………………………………
(vi) Compare whole protein pattern in normal & cancerous cells ………………………………………
(vii) Substituting an amino acid with another at given position ………………………………………
24. Categorize the following protein based products into various categories from a commercial point of view:
(i) Factor VII ………………………………………………………
(ii) Factor IX ………………………………………………………
(iii) OKT-3 ………………………………………………………
(iv) ReoPro ………………………………………………………
(v) t-PA ………………………………………………………
(vi) Humulin ………………………………………………………
(vii) INF alpha ………………………………………………………
(viii) Horse raddish peroxidase ………………………………………………………
(ix) Chymosin ………………………………………………………
(x) kappa casein ………………………………………………………
25. Expand the following abbreviations:
(i) IEF ………………………………………………………
(ii) pI ………………………………………………………
(iii) PITC ………………………………………………………
(iv) HPLC ………………………………………………………
(v) ESI ………………………………………………………
(vi) ELISA ………………………………………………………
(vii) PTFE ………………………………………………………
(viii) LMB ………………………………………………………
(ix) SCHB ………………………………………………………
(x) HBV ………………………………………………………
II Short Answer Questions (2 marks each) Expected time/question: 3- 4 minutes
- Name any four diseases caused due to absence / alteration of key proteins.
- Functional properties of proteins depend upon their 3D structures. Explain with a suitable example.
- Which of the two is a stronger bond and why?
(a) Ionic or covalent bond in vacuum
(b) Ionic or covalent bond in H2O
- When oil is added to H2O, it tends to separate out fairly quickly. Why?
- In proteins hydrophobic regions are preferentially located away from the surface of the molecule and form the interior of the proteins. Why?
- Explain in-situ activation of chymotrypsinogen.
- Factors on which the purification scheme for any given protein depends.
- How does nerve gas result in death of human beings?
- Different between simple electrophoresis & IEF.
- What do you understand by the term molecular pharming? List its advantages.
- List any four industrial uses of the enzyme papain.
- Enlist the various steps needed to maximize protein stability during purification.
- A single E.coli cell produces 20 molecules of a protein called repressor, which has a molecular weight of 20,000 daltons. If the shape of the E.coli cell resembles a cylinder of diameter 1 µm and height 2µm, calculate the number of E.coli cells required as starting material to purify 1 g of the repressor if the purification yield is only 50%.
- Give any four features of large-scale process equipment used for scale up of protein purification.
- How can the number of proteins outnumber the number of genes in the cell?
- The initial load of E.coli in LB broth is 105cells/ml. After incubation at 37ºC for four hours, the load increased to 1015cells/ml. Calculate the specific growth rate and doubling time of E.coli?
- Explain the use of functional non catalytic protein and therapeutic antibodies and enzymes.
- A person suffering from gonorrhoea recovered quickly by constantly having whey protein in his diet. What could be the reason for this?
- How does a charge relay system operate in Chymotrypsin?
- What do you mean by post-transcriptional modifications?
- What is the importance of nutracetical proteins?
- What are zymogens? Where do they get activated?
- Why is sickle cell anemia called a “molecular disease”?
- Why is feeding of colostrum necessary to newborn babies?
- Define nutraceutical proteins. Give two examples.
- What do you understand by the term ‘recombinant vaccines’?
- Give the principle of two-dimensional gel electrophoresis.
- What is the significance of proteins?
III. Short Answer Questions (3 marks each) Expected time/question: 5-8 minutes
1. Different between Sanger’s and Edman’s method of protein sequencing. Which is more beneficial & why?
2. What cause linear proteins to undergo folding in space to form a characteristic shape.
3. Give two examples each of hydrophobic, polar & charged proteins.
4. Differentiate between active & inactive forms of the enzyme chymotrypsin.
5. What happens when glutamic acid is substituted with valine in hemoglobin?
6. Enumerate peptide mapping & list its advantages.
7. Define the following terms: -
(i) GRAS (ii) Downstream processing
(iii) Transgenic animals
8. How can you effectively achieve separation of cellular debris from soluble protein?
9. Depict a generalized downstream process in productive of bulk enzyme preparation from microbial source.
10. List advantages of using a recombinant vaccine based on selected epitopes.
11. Why is it necessary to ensure safety before consuming a genetically engineered food / protein etc.?
12. How can you measure the general effectiveness of a dietary protein source in providing the metabolic requirements of an individual?
13. Study the following enzyme purification table and answer the questions that follow:
Procedure Total protein (mg) Total activity units
step1: crude extract 1000 2000
step2: precipitation {salt} 200 1890
step3: ion exchange chromatography 100 1500
step4: gel chromatography 90 1400
step5: affinity chromatography 2 1000
i] What is the yield of active protein from crude extract?
ii] Write the specific activity for all the 5 steps?
iii] Write the overall yield for all the 5 steps?
iv] Which step in the purification is least effective and why?
v] Which step in the purification is the most effective and why?
14. Explain the possible mechanism of therapeutic uses of whey protein and comment on its general effectives as a dietary protein source.
15. Indicate three important non-covalent forces, which contribute to protein folding and explain how they arise?
16. Describe a method of improving subtilisin.
17. Explain how serine residue in some enzymes becomes reactive. Also suggest how you can confirm that serine residue is involved in the catalysis?
18. What are BCAA? Enlist them? How can BCAA help athletes protect their existing mass?
19. An E. coli cell produces at least 3000 different proteins. One of these is our enzyme of interest produced at a level of 5000 molecules per cell under optimum conditions. If we have to purify 2 gm of this enzyme, estimate how many cells of bacteria will be required theoretically? The molecular weight of the enzymes is 1,00,000 Daltons. Assuming that the cell is a cylinder (d-1µm, h = 2µm). Calculate
(i) The total packed cell volume of E.coli required to produce 1 gm of
intra –cellular enzyme.
(ii) The volume of the fermentor required if the maximum cell concentration inside the fermentor is 2%, as the cells need space to multiply.
20. Explain how protein designing was done in subtilisin.
21. How can sickle cell hemoglobin be identified?
IV Long Answer Questions (5 marks each) Expected time/question:10- 12 minutes
- Name any five methods of characterization of proteins and explain any one in detail.
- Mass spectrometer has emerged as important tool in biotechnology. Justify.
3. Substituting amino acids may change stability of a protein. How? Give one example where such a change has been done in a protein to bring about significant loss of activity.
4. What is the goal of mass spectrometric analysis of biomolecules like
peptides and proteins? Explain two methods, which are used to achieve
this goal. Draw an outline of a mass spectrometer and label the parts and components.
- What is proteomics? Explain what ‘Modern Proteomics’ entails. Why do you think study of this area will be beneficial in future?
- What is protein sequencing? Which was the first protein to be sequenced? Write the chemical reactions involved in the Edman’s sequencing method. Why is it considered an improvement over Sanger’s method?
- Write a short note on the various kinds of non-covalent reactions that take place in the quarternary structure of proteins.
- What is the fate of ions of high molecular mass and low molecular mass in the deflection chamber of a mass spectrometer and how are these ions detected in a detector of a mass spectrometer? Enlist the 4 advantages of using a mass spectrometer.
- What is proteomics? List down and explain different types of proteomics.
- With the help of an example discuss in detail about protein function.
- What is 3D shape of protein? Write in detail about structural organization of proteins.
- (i) Draw a flow chart enumerating the steps involved in the recovery of proteins.
(ii) How is aqueous two-phase partition technique used to isolate protein?
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