14may01final - MCB13U Final Exam, May 14, 2001 R. Schekman...

Info iconThis preview shows pages 1–9. Sign up to view the full content.

View Full Document Right Arrow Icon
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Background image of page 2
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Background image of page 4
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Background image of page 6
Background image of page 7

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Background image of page 8
Background image of page 9
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: MCB13U Final Exam, May 14, 2001 R. Schekman NAME ' (Page 5 cf 13 pages) 7. (35 pts} Yuu have clcned a gene fer a putataive membrane prutein and find the predicted prutein cuntains an Nrterminal signal peptide and three additiunal hydrcphcbic membrane- spanning sequences separated by lung segments cf pular amine acids. A) Given what we learned abcut the signal hypcthesis, draw a diagram predicting the erientatiun cf this prutein after it has becurne assembled in the ER. Include arrcws puintin g tn the cytuplasmic and lumenal surfaces cf the ER, and indicate the pusitiun cf the N- and C- termini cf the prutein. C;- i' 'JW‘HJ .3 Jr ‘i'fi in B) Assume this prutein is transpcrted tn the lyscsume. Draw anuther diagram indicating its erientaticn at the lysuscmal membrane and include an arrew painting to the lumenal surface cf the lysescmal membrane. Indicate the N- and C—termini cf the prutein. F“ C‘W-wflUJ C) Assume as an alternative to part B) that the prctein is tran3purted tn the cell surface. Draw anuther diagram indicating its urientaticn at the cell surface and include arruws painting in the cell estericr and tn the N- and C-termini cf the prutein. FNJJCLFWwUS w 31'! cur-«lac; D) Suppuse ycu have deleted the sequence cf the gene currespundin g tn the secund cf the three membrane anchcr dcmains. Draw twc pictures indicating huw this pretein wuuld be eriented at the ER and at the cell surface {include arruws indicating cytuplasmic. lumenal membrane, and cell surfaces and indicate the N— and C-termini). a . m w. MC3130 Final Exam, May 14, 2001 E L.— \{ R. Schekman NAME “ (Page 6 cf 13 pages) 7. (ccntinuedl E) Suppcse this prctein ccntains the sequence cf N-glycan attachment (asn-x-thn'ser) in each cf its pclar dcmains. Indicate cn diagrams cf the wild type and deleticn mutant prctein (referred tc in part D) (ER and cell surface fcrrns cf each) which pclar dcmains are liker tc acquire N-linked cligcsaccharides (shculd have fcur diagrams tctal). Cal-l ,/ L a.“ H . air “1.11” J. N '- :h DJ I I ' _ (3.1;; {-“fiW-UJ- FL..___ 8. (20 pts} The cystic fibrcsis transpcrter (CF'I'R) 1s a chlcrtde channel. If ycu assume this is the cnly (31' channel in a cell, hcw wcnld ycn expect its absence in cells cultured frcm a patient with cystic fihrcsis tc affect the resting pctentia] cf the cell? Assume that the channel is cpen in cells cultured frcrn a ncrmal perscn. c1 ‘ as pr cut EM kw £2-51” - TF aha PM 11‘ rim/\- .,_-I . U H CNN- abdwmfifilffi), Wav€w_ “dismal rim datum}? am_ W {:Wlfifl Wall taut (L051! ilpfilrcaflfiallmeB Mach il- cklkgffifin can-Juli} L9 . If ycu cculd scmehcw eliminate all Na+ and K+ leak channels frcm a cell that has CFTR, wculd the resulting membrane pctential be negative, pcsitive, or neutral? was, 192mm 0‘ tit-n semi a. canard all. (As-saws; Wand“) Hag {Matrix psmaxfiti'al twvlé‘ Lu. :07 MCBI30 Final Exam, May 14. 2001 R. Schekman NAME ' (Page T of 13 pages) 9. {20 pts} You have created two fusion proteins linking an N-terrninal secretory signal peptide to dihydrofolate reductase (DHFR) and the nuclear localization signal (NLS) to the C- terminus of DHFR. Answer the following about the behavior of these hybrid proteins. A) Will the hybrid proteins be imported into isolated mitochondria, and if so will the reaction occur only during polypeptide synthesis (covtransiational) or after completion of polypeptide synthesis (post-translations? Explain your answer. bwgimia is. .4?” ins. ir‘ia’wt rug—"£31, ltd‘o wtji’sciw_;.g;;3 .laJDJL-E} UR Qfir‘nfljflj W FWIDLC f? l B} Will the cytoplasmic chaperone (hsc'flil) bound to ATP be required for the co— translational translocation of the signal peptide-DEER hybrid protein into isolated ER membrane? Explain. NEH its C®#*Wsl'etxlimal '1 Waggon"! wile anew glam iXW-i (ill—A1 Wino-5W itch Qfilal Ciel-wild: Wain-"M no éb'gliflfiollt (R‘s-{lorithp will Ly +U I”: F; ({er KEEN Chine. iw 34m unfilled Him-$13.1"? ax: C) Will methotrexate inhibit the co~translational transloeation of the signal peptide-DHFR hybrid protein into isolated ER membranes? Explain. qwkoi Fall-“x rll" {mu l fig'k M ppvfigfi ’Tku hHiFéfifiMliflxw-a will irM.§l.enlU we'l’mi'l‘fiii' sac-gal 1 gm 44w 1px“. as». o a C Mimics mg. ' D) Will methotrexate inhibit the import of the DHFR-NLS hybrid into the nucleus? Explain. N E“ gflkw fh’pa.fi$ LN??ij h'rLl‘WlZ'o XWs-WQ 'mzlfo Jr’s: nucleon. “(l/xi lglwb’wz) QC MWb-wmifl' kc: am (All) narrower ml mi w my M03130 Final Exam, May 14, 2001 Y R. Sehekman NAME (Page 8 nf 13 pages) It]. (20 pts} Ynu have cultured mutant fibrnblasts frnm a patient whn has a disease that affects ehelesternl regulatinn. Surprisingly, the mutant fibreblasts will grew nnly if the culture medium eentains free ehelesternl; LDL particles will net replace free ehnlesternl. After a let nf wnrk, but met quite ennugh tn win a Nab-e} Prize, yen diseever that the patient is missing the gene that eneedes the site 1 pretease respnnsihle fer preteelytie maturatinn of the stern] regulatory element binding protein (SREBP). Answer the fellnwing questiens abeut the hehavier ef eells eultured frem this patient. A) Is LDL taken up by reeeptnr-mediateti endneytnsis by these mutant eells? Explain- N e . Prbfimalxstlg M'Eifui‘aiiw UK: 519.15 Mb 'rg Pailuii‘xé 41:: fm&-UQ am "a pit I'M kmamw*p’t‘.m_{a nil-JP. {L}; End’w- hindg i’e «its; 31% W uw—fi—Q VpS'tW'aw. at: law L351. NUS: 0*“ wl'Jnf/xwl’ Aggy 1&ivkif'gfiimlp‘himg’hd’m! nu LbLNLag—JLP xJii'QJ W " millilka - E1 ' *5 LIN... Cw; b4. aw: i Law! w mustard/a}; B) Can these eells make their nwn ehelesterel'? Explain. ERIE E I0 E. saw JYWSMith".m has ate. ENE}; +3 am stag madam... eE ate: \W‘t‘s— Cat: Maths-4.0 app/M . smascm M'ughfi 15 “a . ' when was: amino {in smear—(cc 'hfli‘h‘flmw. Maharth Jttj “st/tam .rcri when {3619? he: Hfl‘fiacafl wué’aa :5 mflq “a. MML. Am mujtewd" call LEW. eh” m'hllu it L3 I C, m g a H .l-I""'- I? C) Are preperly phespherylatetl lysnsnrnal preteins taken up by reeepter—meriiated endneytnsis by these mutant cells? Explain. its. 5269,10 Lt Mk“ was +0 mm at... Mew—abaf west ind-‘31 atoms u-C 3MB P ‘md’iM 'KWSLrw‘f-fitw 4?! an“ shawl-:3 law. he {nélmm MAL-R \eC'EHQX-Rl'iw. Dr Wtaéi’w'é £33193 a? am am reap NEH-2r er“ mug al-r“w~ 934424539 mvaxxeh‘ Bi ARM L‘x’zrhifr'w tit-aw D) Whyr tie these cells requirflelesterel in the grnwth medium? aw- mgww “at BB. Saw-mg Jam. a..er Cam ml: mm LBL. Maggi??? Atf'W/é Q3; txfil‘ {altrung Lhekfi£b>vfli «(m LbL {Mk-wk” ml) 3.4er anfl cm ML malts AM» we“ Ludhflml} Hm W’s-k hem/3 m a Sal-mp cal «CM mubsiml QM gnaw catfish at Ma (JG-J. loci MCB130 Final Exam, May 14, 200] R. Sehekman NAME (Page 9 ef 13 pages) 11. (3D Eta} Previde ene sentence definitiena ef the fellewing: ET R , g A) membrane depelarizatien -- in «#4 WW foliaxd'I-Q'Q f h Quit!“ ’3'“ Qkflcifa EHH dam : 'w; r M (‘a ‘0 La N3 {3* 5 fl“ W M «Mr 5:409 _ fififlflmk B) 't h d'l t' 11 3L2, aka 2:4“ + ’ mine nu ma manxe aperene ._. 1} Fax—re 'w. [w _- m ,riy "Egg,- L- i ‘33“?‘5 it”! “fetid; 1,3 W1. 1: W F ‘ mama. a Wailkmwmi WEN” a gagflw‘ifijgdmeewj -— F “a 1.- 1:! 3 l I I fir-J! C) antipert {Us " if 4 A11 “3H_ ChWMM 1 Mg he? 3"“ “r-E-r‘flw We'th brmgf‘ Hr mm or a.” eC 4.“ GM TR “were um 0Fwa New 5 w WWW M w «m LBW JJLJ'S I [If “E D)pflgfidi ’ ent fangs A W at: limfig a.“ P’Y “ac/“Mme ‘H‘E’i NC” {1-H 3m NEE (nudfl’af {Safe-14' g; f fifla‘iflh 1.5 13. if uf'fi, 1I‘ [-4 mu Clio-1r Pei-«ea a (k f ) Ear: GTPaee activating pretein -. A MR amD away: (Law DJka GTF {aw Shh? wwwam “EaS WWI-i afivmv Iflifii'émt‘iw a?!“ w we; earn; -mfiaw giggfi. m JEN. wwus «Cw U139” w H— H) EOPI] (eeat pretein eemplex H) Liam“ " (373' P '3 1N3 i“ Ag 1 ' 1 e- P‘ W91 e41 "(epl’aa—mk PWrx;}\Qfr~wtg;h4w _' {Wmhlp { ' b1 'Efi MEL] éTflayy$ Preflw “E if” ' ER Mai-34w me- ~ ,3 a I I 1 I 31%- Dfie 'flugenieWfiEvfii; i3 1% fun...“ {j 1 Ah: MT—f It“ vav?(fl2: N3 {1: ATPayji-Fkrai: H I (LU LKM f1? — MI I!” Mics” WW 5‘3 “ME Pm" Ema—Wye; Ira—k LN agar "-. :1) signal reeegnitien particle (SRP) reeepter W 03 why a? firm M Ma whom; aegi- baeag h. a 9.; x Seabfimw 944W aha PRISM {JaLJaPal—gwmcie 3W gun! we: ribosw-t- girl P emw chMflfl‘iiafié-ONQ Mm shins wmw aware; MuUfiLe ‘E‘d‘ WVK JCCJ +1:- Mar Eiffel? sum Lead mm '6'? Mam, //0 MCBIBO Final Exam, May 14, 2001 V Fa- R. Schekman NANIE a" r i (Page 10 of 13 pages) 12. {20 pts) You have isolated a mutation that inactivates the nuclear localization signal {NLS) on the Ran nucleotide exchange factor (GEF). The purified mutant exchange factor has normal activity. Answer the following about this mutant protein. A) Will this mutation cause the depletion or accumulation of Elan-GT1?"I in the nucleus? Explain. ’ m (SUE? meal? is. o tit-La mvcfiqw ’i’o mm ldfl. Q—Ewrfiah€~a A muwrofi shelfxfl “LS. oi: elm he mot... y as? am: '3 W a“; Em C) Will this mutation block or accelerate the rate of cargo protein import into the nucleus? Explain. a“ gmol’sllm at Gwen-"1* ’-~.-. Asa Lg-Ecevolaw‘ .nIH coon carer..- .‘Mfif'firl-fl WU}, of, dit‘temalfij Argus black-Far: CW ihvffi+ rwiko T‘ULiflk-Jf, D) Will this mutation block or accelerate the rate of cargo protein export from the nucleus? Explain. MCBIBEI Final Exam, May 14, 200] k G k/ R. Sehekman NM (Page 1] ef 13 pages) 13. (20 pts} Brefeltlin A (BEA) is a drug that speeifieally bleeks the guanine nueleetide exchange faeter (GEE) that acts en the GTP-binding pretein Arf. In the presence ef BFA, Arf is net reemited te Gelgi er intermediate eempartment membranes and consequently COP! and elathrin ean net assemble en these membranes. A) Describe the eenseqnenee ef BFA treatment ef eells fer seeretery pretein traffie te and threugh the Gelgi apparanls. MCVDLRA Cfif‘i will ie-D EIUCFKQJ}. ER Mam; " ' ' ' . ism. Arm insti'wwfijfiikf we a g} warms-em; as $.41 {mm M mi e {QMBLEQQ‘ Nwl ISMs-EW @ersm 9w“ tent m’c Luz. Wag; t, “q 7) J . m r; , “9 casted. m-Pb-sz Eat-o use W DEDI‘IW'ML d -en Meme Caviar moasz \gvb mefithhq‘ Ki. "Y "3 wet-1* (\LGMM N‘s: Smikfi‘ [EVEN bleefiiggemimx) B) Will BFA treatment bleek the transpert ef Iysesemal preteins frem the trans Gelgi te the endeseme'i' Explain. 5 fl [Siam Q'Akael 914:} U iirlm (“EN' Lie-{’fiitrm cam a.“ lilt'l' \thL/ng tern 5M 2L9 Nth-“gr? Mmfltm'” EM" lfiiflsmxgio He has; thrierewkaxvxg mull mat" FJW~ 3" *(‘i fimfi€flx *‘war - t-t sax-ewe? {Hm Kauai-S“ Hi‘- .31‘ tel-W i015- "i’i—“mi {ND Q WEG‘I'ia C) Will EPA bleek the fermatien ef transpert we ieles frem the ER? Explain W new? CJP‘EI rectbvg ‘Te 94mg,”- ‘NHAUJJ JF-i-afl D) Will EPA bleek the impert and expert ef preteins te and frem the nucleus? Ne +-—"' //,‘2 MCBISD Final Exam, Maj,»r 14, 2001 ;:\J R. Schekman NAME “- (Page 12 pf 13 pages) 14. :20 pts} Tetramethylammcnium icn inhibits the actien cf the vnltage-gated K+ channel. Answer the fellcwing ccncerning the behavinr cf nerve cells espcsed tn this agent. A) Can membranes be depclarized in the presence cf the inhibitnr? Explain. \LO 5. Wing's/mg éLdQDlmmt’BJFiN-“Qfiowrw 2: L] V" H“ " Nil [R'sle QMLQ. if: until I fl“? Uni 1&5] lr+ Llele. t 5 raw: N151 i591" Mahmud N413 DIM ilfilim "‘ slab???- B) Is synaptic transmissicn at a neurcmuscular juncticn blccked if the axcn ccnnected tn the muscle cell is electrically stimulated in the presence cf the inhibiter‘? Explain. Na. Welfi-{Hfifi'lm a “WE/1 hwy “W; {3. 2”; “mg It; and t” “W Ml???” Ca” (WM: tmswms fir:ng Hm: f»th New; .a-'{lu:.2._ts:at. an MJVWH W- News-transmdu lifluw has pflgtggm‘ftm WW Quid/x CEij Mas hamsmmmw 1.63% par h} «\w; W pas‘x’sx-O—wlpl’lc Mfrlmltfl'im‘ C) Can nerve cells be repeater;inr stimulated in the presence ef the inhibitcr‘? Explain. Ne. Kimulfilm. vines stalka Wfllwlfl M19 KCNLLS if adj la lam“ Lal'l’firi ivJ<n\9;\'}l-E’T w'll wrists Lat-lffisfli _ . Malwsxgim my will W MW.“ +5, *5 raglf'msé lpelfiwwjf QCFWSQ up 3?ng \t'l'blli'< dr‘EaEINJS f hclK’ MUSE-Fa”? lit-7' S. l-\ D) Can nerve cells EVETItllfllly he restcred tn a nermal resting pctential if they are electrically stimulated cnce in the presence cf the inhibitcr? Explain. +5 <1) \Ug _.. gigging cf lfi MK Limbs-smgl Ww-Jf‘fiivx‘ wm m5}- //5 MCBUU Final Exam, l‘vIajrr I4, 2001 l,< {j 2.)} R. Schekrnan NAME I (Page 13 cf 13 pages) 15. t 15 pts} Yen have reconstituted the Ca2+-ATPase frcni the sarccplasmic reticulum intc pure phesphelipid vesicles. Answer the fellcwing questiens regarding the prcperties cf these recenstituted vesicles. A) Yen find that the ATPase active site faces the cuter surface cf the vesicles. On which surface cf the vesicle [in the vesicle lumen er en the outside cf the vesicle) will yell intrcduce {Ta2+ in erder ta abserve ATP—dependent active transpert? AL . .+. [PERL—l! m_V-S“‘l" m at- CU“ Eat-{325- g-l: veil-.4 B) Ycu find that the selubilized enzyme in a salutien containing detergent is sensitive te vanadate and ADP. Is lCa2+ transpcrt in the recenstituted vesicle sensitive tc vanadate'll Explain. C) Using different ccnditicns far the recenstitnticn cf the Ca2+ ATPase inte vesicles, yea find that the ATPase activity is cbserved enly when detergent is added to the vesicle preparaticn. Explain this behavior. Use a diagram tc illustrate year ansvver. m h’ffatro \mu la—LQ/m Niecil'flvifi “ML... "uh; he}? in lama-«t Elli! m“ refit '~v’*~W' SW We'd-#71“ Mtge: mfi-‘zsflfig r2; elegarflél malt? ti—am vial-v Wfiri dc? fret—«vth tgctgggplw W ...
View Full Document

Page1 / 9

14may01final - MCB13U Final Exam, May 14, 2001 R. Schekman...

This preview shows document pages 1 - 9. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online