My internet is down due to the storm, and this is the last question I need so I

Question

My internet is down due to the storm, and this is the last question I need so I can submit my assignment tommorow. If anyone can assist me, I would greatly appreciate it. Trying to get this last question done so I can start back up studying for two exams on Monday =(

Answer the following for the two open reading frames (ORFs) given below. (13pts)

Do a BLASTP on the given ORF. The BlastP result is a graphic summary of the conserved domains on the top followed by the distribution of the top 100 Blast hits on 100 subject sequences. Scroll down to the descriptions of the significant alignments. Selecting the specific alignment and sequence ID will give you the answers.

Identify the protein by name, potential function, and name of top match organism, including strain name (if top match says multispecies, select more titles for top organism name).

Give the sequence ID and the Enzyme Commission (EC) number for both proteins.

Name the conserved domains (from the BLASTP results).

Using their EC numbers on the EXPASY site, figure out the protein’s: Optimum pH, temperature stability, secondary structures (number of subunits), localization, storage stability..... http://enzyme.expasy.org/

Ways to access the EC number: Using the Sequence ID link from the Blast result you can scroll down the protein details and locate the EC_number for the protein product.

You can click on the EC number link for more info or enter the EC number in the Expasy enzyme link.

If there’s no EC number in the NCBI protein details, you can search Expasy Enzyme using the protein name.

From the EC page, you can access the various links, Prosite or Brenda to get more detailed information on the protein.

ORF1:

ORF2:

Explanation / Answer

ORF 1 :

1) Identify the protein by name: MULTISPECIES: oxygen-dependent choline dehydrogenase

2) potential function: Choline dehydrogenase - catalyzes the oxidation of choline to betaine aldehyde and betain aldehyde to glycine betaine at the same rate

Choline + acceptor <=> betaine aldehyde + reduced acceptor

3) name of top match organism including strain name : Pseudomonas

4) Give the sequence ID : NCBI Reference Sequence: WP_003104848.1

5) the Enzyme Commission (EC) number : 1.1.99.1

6) Name the conserved domains from the BLASTP results: PRK02106

EXPASY site information:

8) protein’s optimum pH: 9  

9) pH stability: 8-9.5

10) temperature stability: 35 -50

11) secondary structures (number of subunits): 1, it is a monomer

12) localization: a) Enzyme is found on the membrane as peripheral membrane protein.

b) It is also seen particle bound

13) storage stability: -200C , stable.

ORF 2:

1. sequence ID:  1X7N_A

2. Identify the protein by name: Chain A, The crystal structure of Pyrococcus furiosus phosphoglucose isomerase with bound 5-phospho-D-arabinonate and Manganese

3. potential function: protein similar to glucose-6-phosphate isomerase, catalyzes the reversible isomerization of glucose-6-phosphate and fructose-6-phosphate

4. name of top match organism including strain name: Pyrococcus furiosus

5. Enzyme Commission (EC) number for protein:

6. Name the conserved domains (from the BLASTP results) : PRK04190

7. EC number: 5.3.1.9

on the EXPASY site:

8. alternative names:

9. Protein’s optimum pH: 7

10. temperature stability: 85

11. secondary structures (number of subunits): 2, it is a homodimer

12. localization: not given for this particular organism

13. storage stability: not given for this organism

Related Questions

Navigate

• Browse (All)
• Browse M
• Subjects

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.