Exam 1 from 2008 (with answers) - BMB/MICRB 460 / February...

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BMB/MICRB 460 / February 14, 2008 NAME: Exam 1 / TEST FORM A 1 TEST DIRECTIONS: Choose one answer for each multiple-choice question and RECORD it on the line to the left of the question. Write your answer to the short answer questions LEGIBLY in the space provided Each question is worth 4 points unless otherwise indicated. I will post the exam, the answers and scores as soon as all students have taken the exam and grading is complete. 1 . (6 pts) Draw or describe how Ran exists in two different states to act as a “switch” protein and create a different environment in the nucleus versus the cytoplasm. Include discussion of 1) specific regulatory proteins necessary for switching between states 2) the mechanism for switching between states and 3) note which state predominates in the nucleus. Nucleus RanGTP predominates due to action of guanine nucleotide exchange factor (GEF) RCC1 which causes Ran to let go of bound GDP and bind to GTP (nucleotide exchange) Cytoplasm RanGDP predominates due to action of GTPase activating protein (GAP) called Rna1p and its accessory protein RanBP1 which activate the intrinsic GTPase activity of Ran causing it to cleave/ hydrolyze the gamma phosphate from GTP and leaving Ran in the GDP bound form This allows Ran to act as a switch because it can bind some proteins (importin beta) in the GTP bound form but not in the GDP bound form. 2 . I experimented to figure out why vSNAREs were restricted in the Hu paper while they were not restricted in the Weber paper. Take a look at the results below and choose the appropriate conclusion. Lane 1= soluble tSNARE + complex proteoliposomes (PE+PS+PC+cholesterol) w/ recombinant vSNARE Lane 2= soluble tSNARE + simple proteoliposomes (PE+PS+PC) w/ recombinant vSNARE Lane 3= soluble tSNARE + complex proteoliposomes (PE+PS+PC+cholesterol) w/ native vSNARE Lane 4= soluble tSNARE + simple proteoliposomes (PE+PS+PC) w/ native vSNARE A. Lipid is causing restriction of the vSNARE B. The tSNARE causes the restriction of the vSNARE C. The vSNARE purified from cow brain is restricted while the bacterially produced vSNARE is not D. Native vSNARE and cholesterol are required to restrict the vSNARE 1 2 3 4 SDS PAGE Coomassie stained
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BMB/MICRB 460 / February 14, 2008 NAME: 2 3. To examine the subcellular localization of VIP (Very Interesting Protein) in a cultured cell system in your lab, you need an anti-VIP antibody. In which animal would you choose to make your primary antibody so that you could use the fluor-conjugated goat anti-rabbit secondary antibody in your lab freezer that works well for indirect immunofluorescence? A)
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This note was uploaded on 10/19/2009 for the course B M B 460 taught by Professor Hanna-rose,w during the Spring '08 term at Pennsylvania State University, University Park.

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Exam 1 from 2008 (with answers) - BMB/MICRB 460 / February...

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