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13 Dec 2019

What would be the affect of the following conditions on the respective biosignaling pathway we discussed in class? (Biochemistry II)

a. The presence of the following GTP analog.

b. A mutated SH2 domain.

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Related questions

You are studying the role of dynamin in the budding and pinching off mechanism of clathrin coated vesicles from the plasma membrane. You know that dynamin uses GTP hydrolysis to perform its function, but you would like to test how dynamin behaves in the presence of a non-hydrolyzable analog of GTP. Describe an experiment that allows you to observe the function of dynamin in the presence of either GTP or a non-hyrolyzable GTP analog. What do you predict will happen in the presence of the analog (Be detailed in your response)?

1) how does the timing and location of Arf1-GTP hydrolysis contribute the transport of COPI coated vesicles?

2) Consider cargo proteins that are destined for secretion to the extracellular space. For each of the following pairs of mutations or conditions, predict where the cargo will end up (ex. in the cytoplasm, a specific organelle, transport vesicles, multiple places, extracellular space) for: (i) & (ii) Each individual condition; & (iii) The combination of both.

A.(i) Normal Sar1 is mutated so that it hydrolyzes GTP immediately after binding GTP.

(ii)The signal recognition particle is mutated to be unable to bind to the SRP receptor.

(iii)Both


B.(i) A mitochondrial signal sequence is added to the C-terminus of the cargo protein.


(ii)v-SNAREs on COPII coated vesicles are mutated to be unable to bind t-SNAREs


(iii)Both


C. (i) COPII coat proteins are mutated so they don’t bind to the ER membrane.


(ii)COPI coat proteins are mutated so they don’t bind to the Golgi membrane.


(iii)Both


D.(i) Normal Sar1 is mutated so it binds tightly to GDP and cannot exchange it for GTP. (This leads to Sar1*-GDP being the only form of the protein in the cell.)

(ii) A transmembrane domain is added to the cargo protein.

(iii)Both

The outer segments of rod photoreceptor cells can be broken off, isolated, and used to study the effects of small molecules on visual transduction because the broken end of each segment remains unsealed. How would you expect the signaling pathway to be affected following the addition of a non hydrolyzable analog of GTP?

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