ࡱ> Root Entry'"@(FileHeaderlDocInfoZBodyText @` !#$%&'()*+,-/10.23456Root Entry'@(FileHeaderlDocInfoZBodyText @` 78] <\><Proteomic architecture mapping by proximity labeling> <\Դ><Recently, proximity labeling has been developed to map spatially localized proteome in live cells. Usually, these methods employ enzymatic biotinylation of the proximal proteins with short-lived reactive biotin species. The labeled proteins may contain biotinylated modifications, which can be detected by mass spectrometry after enrichment by streptavidin beads through affinity purification. Our lab has developed the direct detection method of biotinylated modifications on the labeled proteome (Spot-ID) and using this method, we could reveal the in vivo topological direction of 135 inner-mitochondrial membrane (IMM) proteins by an in situ-generated radical probe with genetically targeted peroxidase (APEX) in live cells. Furthermore, we could identify the proteomic architecture of outer mitochondrial membrane (OMM) proteome by using isotope-coded labeling in a live cell. 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ĿCbTMr{x.5"bM8{y3#2a4!uW^w⣜I}@S`:tmS bf PX3\=)M`MAhQ͡wݘo~ӉT!yE童|juљ GhҹkڐӚÚDn:J LMŁ(GC3E憑*v_mD;oy׭q1Jm,[ܢ%pY^ D&$uP9w?|,ӻmL+lE^ɎYR9;QIZI&?.4˧J2Ods%R¿X/`kAǿ;ylAAV3ET` X)&` ăCR=(Ģxi̹`sA{gE5{_yf"}/Xt1 (![IHLɮN2Q=NX PٚHDsP6Ia /SR<Žq$vNqI;i`'Wa v$ h#>f 9a <IU]l,yB HJ)CpZ[]gJ瀔$Nĵ\&pbIU6<+%&0("{>5If⏳3$i^/+4YخvZ 󸁡zTXW(zYGa ܧZcn= p;fUF۔oVpINW[+XFՙqJ2#G(8~rKwk=9n'ȍO_?Ǜ%ʛ( ֙FjvK+*FnWŪHWP Document FileJ\*?/,9bwweMKu@DO 6C,H,#GB!k0~ 6*7=M*Z:ҙ#?*?ӿT@Y~X+mj3'm[v|:j)%C$Mϼ5#cd߈rgeted peroxidase (APEX) in live cells. Furthermore, we could identify the proteomic architecture of outer mitochondrial membrane (OMM) proteome by using isotope-coded labeling in a live cell. Using this method, we could successfully determine the membrane>WOhe3$[@ 89V$MD J@N&مݝegv ͡o<0AAz5}Bt/ K]e%YMۨ3L\}ݐڐNrwlFj/'PIrm r yc_PG0ێ- 5  Z4 hP$i@GeIS@WD!=xSUipVd ,`MYrC/I;pSk݁:J<>tJ|OG%i@ݕru%ŀV%>!k-:gf]T*OT×.6|] Я6*k{zÚx`=S0{Fl-B#e6#!ߖ̪b@fU)7`h#ߙ+ΫbQgu&X8^y^8oK#Py:$&eowՒR弶+Fe"*!ΒTJfHzjUTV)a*HPE'o+R>*V(+|HZ sCL-jg\ݥ;F3MޛE{Ipy5 Û4 ;' Ha"un݀EёT6܃U jalS2^NaQh 3#)Vg_霶T>L](>wvp Ͷj6 b+;3/hV|OڬbMErmj6.RghiMPgu.]~ׅ)u* lCcJ\:ltAܣ**i.2*gXg ,SKm\Utx]b4펾Zq>UN&9 wA<- Ou4h^;YSqV3"/X{OWs&YQм1wnF}p < 䋑?߮C|8{n8[}Gf=7eL"Ogy~<[L:S:g0_x%['|BW-elg!+SVl!k׈VkA~&iIŢB"&.?Z4!ֶPLVŃA&b=x=zP=dfltɾy;{߼yy/QmCtO%~f>:1)װ=*Bܫþ?ʁJ<50|`vM]SA~ycmkZt I'tJ:~)"*J=vWj*ᴘ@]?@ABCDEFGHIJKLMNOPQSTUVWXY[\]^_`abcdefghijkpmnorsvwxyz{|} !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQSTUVWXY[\]^_`abcdefghijkpmnorsvwxyz{|}HwpSummaryInformation.RPrvImage [ PrvTextDocOptions Section0~Scripts 'JScriptVersion t DefaultJScriptq_LinkDocu 8] <\><Proteomic architecture mapping by proximity labeling> <\Դ><Recently, proximity labeling has been developed to map spatially localized proteome in live cells. Usually, these methods employ enzymatic biotinylation of the proximal proteins with short-lived reactive biotin species. The labeled proteins may contain biotinylated modifications, which can be detected by mass spectrometry after enrichment by streptavidin beads through affinity purification. Our lab has developed the direct detection method of biotinylated modifications on the labeled proteome (Spot-ID) and using this method, we could reveal the in vivo topological direction of 135 inner-mitochondrial membrane (IMM) proteins by an in situ-generated radical probe with genetically targeted peroxidase (APEX) in live cells. Furthermore, we could identify the proteomic architecture of outer mitochondrial membrane (OMM) proteome by using isotope-coded labeling in a live cell. 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ĿCbTMr{x.5"bM8{y3#2a4!uW^w⣜I}@S`:tmS bf PX3\=)M`MAhQ͡wݘo~ӉT!yE童|juљ GhҔJ\*?/,9bwweMKu@DO 6C,H,#GB!k0~ 6*7=M*Z:ҙ#?*?ӿT@Y~X+mj3'm[v|:j)%C$Mϼ5#cd߈rgeted peroxidase (APEX) in live cells. Furthermore, we could identify the proteomic architectkڐӚÚDn:J LMŁ(GC3E憑*v_mD;oy׭q1Jm,[ܢ%pY^ D&$uP9w?|,ӻmL+lE^ɎYR9;QIZI&?.4˧J2Ods%R¿X/`kAǿ;ylAAV3ET` X)&` ăCR=(Ģxi̹`sA{gE5{_yf"}/Xt1 (![IHLɮN2Q=NX PٚHDsP6Ia /SR<Žq$vNqI;i`'Wa v$ h#>f 9a <IU]l,yB HJ)CpZ[]gJ瀔$Nĵ\&pbIU6<+%&0("{>5If⏳3$i^/+4YخvZ 󸁡zTXW(zYGa ܧZcn= p;fUF۔oVpINW[+XFՙqJ2#G(8~rKwk=9n'ȍO_?Ǜ%ʛ( ֙FjvK+*FnWŪHWP Document Fileure of outer mitochondrial membrane (OMM) proteome by using isotope-coded labeling in a live cell. Using this method, we could successfully determine the membrane>WOhe3$[@ 89V$MD J@N&مݝegv ͡o<0AAz5}Bt/ K]e%YMۨ3L\}ݐڐNrwlFj/'PIrm r yc_PG0ێ- 5  Z4 hP$i@GeIS@WD!=xSUipVd ,`MYrC/I;pSk݁:J<>tJ|OG%i@ݕru%ŀV%>!k-:gf]T*OT×.6|] Я6*k{zÚx`=S0{Fl-B#e6#!ߖ̪b@fU)7`h#ߙ+ΫbQgu&X8^y^8oK#Py:$&eowՒR弶+Fe"*!ΒTJfHzjUTV)a*HPE'o+R>*V(+|HZ sCL-jg\ݥ;F3MޛE{Ipy5 Û4 ;' Ha"un݀EёT6܃U jalS2^NaQh 3#)Vg_霶T>L](>wvp Ͷj6 b+;3/hV|OڬbMErmj6.RghiMPgu.]~ׅ)u* lCcJ\:ltAܣ**i.2*gXg ,SKm\Utx]b4펾Zq>UN&9 wA<- Ou4h^;YSqV3"/X{OWs&YQм1wnF}p < 䋑?߮C|8{n8[}Gf=7eL"Ogy~<[L:S:g0_x%['|BW-elg!+SVl!k׈VkA~&iIŢB"&.?Z4!ֶPLVŃA&b=x=zP=dfltɾy;{߼yy/QmCtO%~f>:1)װ=*Bܫþ?ʁJ<50|`vM]SA~ycmkZt I'tJ:~)"*J=vWj*ᴘ@]