Structure and kinetic characterization of human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, GAPDS Apirat Chaikuad, Naeem Shafqat, Ruby Al-Mokhtar, Gus Cameron, Anthony R Clarke, Leo Brady, Udo Oppermann, Jan Frayne, Wyatt W. Yue To cite this version: Apirat Chaikuad, Naeem Shafqat, Ruby Al-Mokhtar, Gus Cameron, Anthony R Clarke, et al.. Structure and kinetic characterization of human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, GAPDS. Biochemical Journal, Portland Press, 2011, 435 (2), pp.401-409. . HAL Id: hal-00581535 https://hal.archives-ouvertes.fr/hal-00581535 Submitted on 31 Mar 2011 HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 )'*)*'$! $) ')' /) %$%*#$(&'#3(& ".'".3 ;3&%(&).'%$(1 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t +$-/#$&0KV> (#!,/KV>04'E*&#/-LV>0.( -*)L>)/#*)4A'-& L>A * -4L>*++ -())KM>)-4) LW)4//A0 KW K /-0/0-' )*($.*).*-/$0( )$1 -.$/4*!3!*- 3!*->X\ L #**'*!$*# ($./-4 )$1 -.$/4*!-$./*' -$./*'>]V M */)- . -# )/- $*( $' . -#)$/ 3!*->X\ V # . 0/#*-.*)/-$0/ ,0''4/*/#$.2*-&A W *-- .+*) ) (4 - .. /* $/# -*!/# . 0/#*-.G ($'%)A!-4) Q-$./*'AA0&*- 24//A40 Q."A*3AA0&HA #*-//$/' @/-0/0- )&$) /$.*!#0() V Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t 0().+ -(E.+ $!$"'4 -' #4 EXE+#*.+#/ #4-*" ). G#H$."'4*'4/$ )54( .. )/$'!*-/# .0-1$1'*!.+ -(/*5*>)*)./$/0/ .+*/ )/$'/-" /!*-)*)E#*-(*)' *)/- +/$*)A *2 1 -> )54( #-/ -$5/$*)*!#. )#(+ - 4/# $!!$0'/4$) +-*0$)".*'0' - *($))/+-*/ $)A - >2 #1 *1 -E 3+- .. $))#$"#'4E.*'0' !*-(*! #/-0)/ //# E/ -($)0.G#eH>)-4./''$5 /# #*(*E/ /-( -$ )54( $)/2* '$")*(+' 3 .A# #eEbE+#*.+#/ ./-0/0- (+./# /2*)$*)- *")$/$*).$/ .2$/#$) /# /'4/$+*& //#/*-- .+*)/*/# *). -1 ..$/ )/# ) 2'4- *")$. $.$/ $ )/$!$ $)*/# -*-")$.(.A# #eEbE"'4 -*'./-0/0- .#*2.. - )$+$/*0.$)$)"*!"'4 -*'/ /# .)) 2$.$/ .> (*)./-/$)"/# +-*+ ).$/4*!/# . )$*)- *")$/$*).$/ ./*$))*)E +#4.$*'*"$''4E- ' 1)/'$").A*(+-$.*)*!&$) /$+-*!$' . /2 )#e)$/..*(/$ ,0$1' )/- 1 '.(* -/ XE!*'$)- . $)/'4/$ !!$$ )4!*-#eA#$.(4 //-$0/' /*.0/' ($)*$.0./$/0/$*).+ -$+# -'/*/# /$1 )/- /#/$)!'0 ) /# #-" +-*+ -/$ .)+-*/*)/$*).// .*!/'4/$- .$0 .A0-//# - !*- '0$/ ./-0/0-') &$) /$! /0- .*!#/#/($"#/+-*1$ $).$"#/!0'$)!*-(/$*)/*2-.$)#$$/*- 1 '*+( )/A '+ ) %$( >"'4 -*' #4 EXE+#*.+#/ #4-*" ). ?>"'4 -' #4 EXE+#*.+#/ #4-*" ). >.+ -(E.+ $!$?X>"'4 -' #4 EXE+#*.+#/ ?>+-*/ $)/)&?>-**/E ( )E.,0- 1$/$*)A W Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce *(+- /*.*(/$ .2#$##1 )./-0/0-''4)&$) /$''4#-/ -$5 G- 1$ 2 $) IW]JH>/# .+ -(E.+ $!$$.*54( #.+-*1 )$)/-/' /*./04'-" '40 /*/# $).*'0$'$/4*! )/$1 )- *($))/+-*/ $).A )/'4>/# ./-0/0- *!# / -*E/ /-( -$*(+' 3*!-/E ) #. )- +*-/ /*(* -/ - .*'0/$*)>IW^J>)+-*1$ /# !$-./1$ 2*! ((('$)>' $/2$/#$))0))/0-'*'$"*( -A - >2 #1 *1 -E 3+- .. #0() /-0)/ //# E/ -($)0.G#eH>/*"$)$).$"#/.$)/*$/../-0/0-')&$) /$+-*+ -/$ .A# #e#*(*E/ /-( -$-4./'./-0/0- .> / -($) $)*(+' 3 .2$/#+#*.+#/ )b G#eEbEYH)2$/#"'4 -*')bG#eEbE*'H>+-*1$ /# !$-./#$"#E Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t # "'4*'4/$ )54( "'4 -' #4 EXE+#*.+#/ #4-*" ). G ?AAVAWAVAVWH/'45 ./# bE + ) )/*3$/$1 +#*.+#*-4'/$*)*!E"'4 -' #4 EXE+#*.+#/ GEXH/*!*-(V>XE $+#*.+#*"'4 -/ GHA $.2$ '4- *")$. .-0"/-" /0 /*$/. )/-'-*' $) "'4*'4.$.>)$))*)E"'4*'4/$+-* .. ..0#.)0' -/-).+*-/>- +'$/$*))- +$-> ( (-) !0.$*))+*+/*.$.IVJA*- 3(+' > .!-*(/-4+)*.*(/$.+ $ .>2#$# + ) # 1$'4*)"'4*'4.$.!*-+-*0/$*)>- +-*($.$)"-0"/-" /./**(//-*+$'+-.$/$$. . . IWEYJA)((('.>0(0'/$*)*! $)/# )0' 0.$.'$)& /* '' /#IZJ># ) $/.$)#$$/$*) ($"#/+-*1$ ))/$E+*+/*/$++'$/$*)A). 1 -') 0-* " ) -/$1 $. . ...*$/ 2$/#/# 3+- ..$*)*!(0/)/+*'4E"'0/($) +-*/ $).> A"A 0)/$)"/*)C.$. . G H> #. ).#*2) /*$)/# (0/)/+-*/ $).I[JA '.*- +*-/ '4$)./# (4'*$+- 0-.*-+-*/ $)GH> $(+'4$)"+*/ )/$'-*' $)/# 1 '*+( )/*!'5# $( -O.$. . I\JA (('.+*.. ..)*/*)'4/# .*(/$ $.*54( +- . )/$)''/$..0 . 3 +/.+ -(/*5*>0/ '.*.+ -(E.+ $!$$.*54( GH 3+- .. /'/ ./" .*!.+ -(/*" ) .$.I]>^JA.#*2. .$")$!$)/. ,0 ) $1 -" ) !-*($/..*(/$*0)/ -+-/A*- 3(+' >#0()G#H .#- .*)'4Z[a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bA# - .0'/$)"/#$*4'E )54( $./# )//& 4)$)*-")$ +#*.+#/ /*!*-(A)/#$.(* '>/# X+#*.+#/ *!.0./-/ EX$. 3+ / /*$)/*/# $ .$/ 0-$)"/# !*-(/$*)*!/# # ($ /'$)/ -( $/ >)!'$+./*/# ..$/ !*- #4-$ /-).! -> /*1/ /# $.$/ !*-)$)*($)"$)*-")$+#*.+#/ IWU>W\JA X Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 - .*'0/$*)1$ 2*!/# /2*)$*)- *")$/$*).$/ .$)#0() )54( A*($) 2$/#*(+-/$1 &$) /$)'4.$.>/# . ./-0/0- .+-*1$ -$"*-*0. .-$+/$*)*!/# .+ -(E.+ $!$$.*54( /#/(4 ..$./$)#$$/*- .$")A Ac ce pte d Ma nu THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 sc rip t Y Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce )'*)*')'# $) %$ */#-4./'!*-(. '*)"/*/# E )/ - (*)*'$)$.+ "-*0+WG' VHA/-0/0- .2 - .*'1 4(*' 0'-- +' ( )/2$/# IXVJ0.$)"/# -/.+ -(**-$)/ .IW^J.. -# (* 'A ).$/4(*$!$/$*))1 -"$)"2 - + -!*-( 2$/#IXWJ>)/# $(+-*1 +#. . 2 - 0. !*-0/*(/ (* '0$'$)"2$/#0) -IXXJA/ -/$1 4' .*!()0'(* '0$'$)" 0.$)"IXYJ'/ -)/ 2$/#- !$) ( )/$)ZIXZJ2 - + -!*-( A//# './- !$) ( )/ ./ +- ./-$) - !$) ( )/2.++'$ 0.$)"/ ).*-"-*0+. / -($) 4/# . -1 - IX[JA/*'' /$*))- !$) ( )/.//$./$.- .0((-$. $)' VA ).3())$/.#! $) ( *($))/#e2.$'45 "$)././*-" 0!! -*)/$)$)"/$1/ #-*'/*- (*1 *0)*!/*-> )'$)"0-/ ( .0- ( )/*!+-*/ $)*) )/-/$*)A3($)/$*)*!/# +-*/ $) .+ /-.#*2 )*b.*-) /W[U)(.#*2$)"/#//# b# ).0 ..!0''4- (*1 A $) /$..4.2 - . *)/# !*-2-- /$*)>$)2#$#+#*.+#*-4'/$*)*!EX2./'45 $) /# +- . ) *!b)$)*-")$+#*.+#/ IX\JA''- /$*).2 - + -!*-( $)VU(.*$0( +4-*+#*.+#/ >WU(.*$0(+#*.+#/ + ]AZ>Xf$/#$*/#- $/*')VU(.*$0(-. )/ > 0.$)"ZUU)"D(' )54( A# !*-EX2.( .0- 0.$)"*) )/-/$*)-)" UAXEYAU(/ !$3 b*) )/-/$*)GUAZ(HA# !*-b2. / -($) 0.$)"*) )/-/$*)-)" UAUWEV (/!$3 EX*) )/-/$*)GVAZ(HA''- " )/.+-/!-*(#e2 - $.+ ). $)^[E Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t '%) $-&'(( %$$&*' ) %$ !-"( )/ )*$)"#G#e>[^EX^]? ))& )/-4\[Z\VV[H2..0'*) $)/* /# +E 1 /*-G ))& )/-4VWYUVZU\^H$)*-+*-/$)"E/ -($)'E' 1' $.[E/"A # *-- .+*)$)"+'.($2./-).!*-( $)/*"!WVGXHEX>0'/0- $)V*! --$!$ -*/#/X\R>)$)0 2$/#UAZ(*1 -)$"#//V]RA ''.2 - #*(*" )$5 $)'4.$.0!! - GZU( + \AZ>ZUU('>ZaG%D%H"'4 -*'>WU($($5*' H)$).*'0' (/ -$'2. - (*1 4 )/-$!0"/$*)A# .0+ -)/)/2.+0-$!$ 4!!$)$/4G$E. +#-*. H).$5 E 3'0.$*) G0+ - 3\ZH#-*(/*"-+#4A0-$!$ +-*/ $)2./- / 2$/# $.E/"" +-*/ . *1 -)$"#// YR>)/# )+.. *1 -V(*!$E +#-*. - .$)A# E/- / +-*/ $)>*)/$)$)"/# - ())/ 1 /*-E )* . ,0 ) BC//# E/ -($)0.>2.*) )/-/ /*VW("D(')./*- $) ./*-" 0!! -GVU( + \AZ>ZUU('>ZaG%D%H"'4 -*'>UAZ(/-$.GWE -*34 /#4'H+#*.+#$) #4-*#'*-$ GH/E]URA '.()"" /) %$$)%"") %$ #e2.-4./''$5 /YR$)VZUE)'-*+.0.$)"/# .$//$)"-*+1+*0-$!!0.$*)( /#*A$' -4./'.*!/# #eEbEY*(+' 32 - */$) 4($3$)"+-*/ $)+- E$)0/ 2$/#Z( bG$"(H2$/#/# - . -1*$-.*'0/$*)GWUaXXZU>UAWD+#*.+#/ )VUaG%D%H /#4' ) "'4*'H$)V@W1*'0( -/$*A-4./'.*!/# #eEbE*'*(+' 32 - !*-/0$/*0.'4*/$) !-*(($3$)"+-*/ $)'*) 2$/#- . -1*$-.*'0/$*)GWUaXXZU>UAWWY>VUaG%D%H /#4' ) "'4*')UAV$.E/-$.+-*+) + [AZH$)V@V1*'0( -/$*A// (+/./*-4./'$5 #e$)/# !*-(#1 )*/ ).0 ..!0'/*/ A''-4./'.2 - -4*E+-*/ / 2$/#- . -1*$-.*'0/$*) .0++' ( )/ 2$/#WZaG1D%H"'4 -*'>)!'.#E**' $)'$,0$)$/-*" )A$!!-/$*)/2 - *'' / *) ('$) UX//# $(*)$"#/*0- G#eEbEYH)$)E#*0. *)$"&0 E0+ --$"#/.*0- G#eEbE*'HA/2 - +-* .. 2$/#)!-*(/# Y0$/ IXUJA Z Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip t 2 ''+'/ >)+- E$)0/ /WZR!*-W($)A# - /$*)2.$)$/$/ 4/# $/$*)*!#eA / 4E.// - /$*).2 - - *- /XYU)(!*-X($)2$/#- $)"./& ) 1 -4W. >0.$)" (3($-*+'/ - -G*' 0'- 1$ .*-+>HA# /)" )///$( 5 -*2.2 '' !$) )0-/ '4 !$) /# $)$/$'1 '*$/4A'0 .!*->(3)/2 - '0'/ 0.$)"-+# -$.(G-+#*!/2- )AH4)*)E'$) -- "- ..$*))'4.$.A# .( +-* ..)+-'' '&$) /$ ..4.2 - '.*+ -!*-( 2$/##0() !-*( -4/#-*4/ .G$"(HA (( %$$*#'( /*($**-$)/ .)./-0/0- !/*-.#1 ) +*.$/ $)/# -*/ $)/)&0) -/# ..$*))0( -.X#^ G#eEbEYH)X+!2G#eEbE*'HA Ac ce pte d Ma nu THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 [ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t %# $$)&'%*) %$$()'*)*')'# $) %$%C *$ )/$!4/# *+/$('*($)*0)-4*!#!*-./-0/0-'./0$ .>2 3+- .. . -$ .*!!0''E ' )"/#)/-0)/ *)./-0/.$)).DE/ -($)''4 $.E/"" +-*/ $).G$"0- VHA *. -1 /#//# !0''E' )"/#+-*/ $)2.+- *($))/'4$).*'0' 2# )- *($))/'4 3+- .. A $"# ' 1 '.*!.*'0' 3+- ..$*)2 - !*0)$). 1 -'E/ -($)''4/-0)/ *)./-0/.#-*0-$)"/# $.E /"//# E/ -($)0.>2#$' !0.$)"/# !!$)$/4/"/*/# E/ -($)0.*!/# ,0$1' )/*)./-0/.- ) - /# (' ...*'0' A .0. ,0 )/'4+0-$!$ /# E/ -($)' $.E/"" *)./-0/ )*(+..$)"[^E YU\G- ! -- # - !/ -.#eH/*#*(*" ) $/4G$"0- VH)$/.-4./'./-0/0- 2. / -($) $)/2**(+' 3 .@*) *0)2$/#b)$)*-")$+#*.+#/ G#eEbEYH/ VA\W- .*'0/$*)>)/# */# -*0)2$/#b)"'4 -*'G#eEbE*'H/WAVZ - .*'0/$*)G' VHA )*/##e*(+' 3 .>/# .4(( /-$0)$/*(+-$. ./2*.00)$/.)G8E /2 ) .00)$/.NUAVEUAWHG$"0- VH>2#$#" ) -/ )*/# -.00)$/+$-)4/2*!*'.4(( /-4 *+ -/$*)G'4$)"++-*3$(/ '4*)/# +') >0.$)"+- 1$*0.)*( )'/0- IX]JH>- .0'/$)"$) #*(*/ /-( -2$/#WWW+*$)/.4(( /-4G$"0- VHA$($'-/**/# - .> #.00)$/*).$./.*! )E/ -($)')0' */$ $)$)"*($)GE*($)?[^EWWUH)E/ -($)'/'4/$*($)GE *($)?WWXEYU\HA# E*($)*).$./.*!) $"#/E./-) 9E*- !')& 4# '$ .*)*/# .$ .$)'..$'*..())!*'A# E*($)$.0$'/!-*(!$1 E./-) 9E.# /*- 2$/## '$ . *)*) .$ >)#-*0-./# /$1 E.$/ /'4/$- .$0 .4.WWY) $.WZV>.2 ''.*). -1 E .#+ '**+GE'**+?WZWEW\]H/#/!*-(./# / /-( -$*- A# E'**+*)./$/0/ ./# ($).$/ *! $)/ -.00)$/*)//.)#.+- 1$*0.'4 ).0"" ./ /* / -($) D*!/*-.+ $!$$/4 IX^JA# #e./-0/0- .- .$($'-/**/# - . A"A#0()'$1 -D+' )/'G# HIWWJ> $""#!#! $"G" HIWU>W\>YUJ>)G HIV^J) "& G HIWEYJ )54( .>. 1$ ) 48E1'0 .-)"$)"!-*(UA\EWAX' $/.#-$)"*)'4 WUE[\a. ,0 ) $ )/$/4A C%#%3))'#' # ,0/ -)-4--)" ( )/)$)/ -.00)$/$)/ -! .*!#e- *).$./ )/2$/#/# !0)/$*)' *'$"*( -$.// +-*+*. !*-*/# - .>2$/#.0/' $!! - ) .$)/2*! /0- .)( '4/# ' /-*.//$.0-! +*/ )/$')$)/ -.00)$/. ' /$1$/4' !/A# #e./-0/0- .- 1 '/#- +-*($) )/'4#-" +/# .//# $( -$)/ /-( -$$)/ -! .>*)/-$0/ !-*(/# +*.$/$1 '4E #-" E'**+GWZWEW\]H>/# ) "/$1 '4E#-" XZUEX\W- "$*))/# ($3E#-" VU[EVXU - "$*)G$"0- VHA# 2$ .+- #-" $./-$0/$*)$)#e$.(*- +-*($) )//#).*(/$ # G$V0]!>V5),H0 /*. 1 -')*)E*). -1 #-" D+*'-- .$0 .G A"A'0VVW>4.VV^> 4-VW\>4.VW]> -WZW>4-WZX>-"W[Z> $.W\Z>.+XZV>'0XZW>4.X\UHG$"0- VH>).0"" ././#/ #e/ /-( -$5/$*)$.!$'$// 1$$)/ -.00)$/ ' /-*.//$*(+' ( )/-$/4A*- 3(+' >/ /# $( -$$)/ -! /# +*.$/$1 '4E#-" E'**+*!.00)$/+-*/-0 .$)/*) "/$1 '4E#-" "-**1 !-*(/# XZUEX\W- "$*)*!.00)$/>- /$)")0( -*!.'/-$" .G$"0- WHA//# / /-( -$$)/ -! >'*. /*/# *!/*-$)$)".$/ >/# ($3E#-" VU[EVXU- "$*)*!.00)$/ +&."$).//# +*.$/$1 '4E#-" E'**+*!.00)$/)/# ) "/$1 '4E#-" XZUEX\W- "$*)*! .00)$/ . *)$!! - )/$/$)"! /0- /2 )# )#e$./# . ' /$1$/4' !/% )//*/# )*.$) +*& /*!/# bE$)$)".$/ G$"0- VHA# . ' /$1$/4' !/ 3#$$/.-*..E.+ $ . \ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t 1-$$'$/4$).$5 >)#. ) 3+'*$/ !*-.(''E(*' 0' $)#$$/*- 1 '*+( )/. 3 (+'$!$ 4 /# . ' /$1 $)$)"*!) )*.$) -$1/$1 I[EGVE)+#/#' ) ( /#4'HEW′E *34EW′EGX>ZE $( /#*34 )5($*H )*.$) J/*/# . ' /$1$/4' !/*!/# /-4+)*.*(/$ G$V$XWH IWEYJA)*0-#e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e. ' /$1$/4' !/A0-/# -(*- >/# -"W[Z.$ E #$)!*-(. ' /-*.//$$)/ -/$*).)2/ -E( $/ #4-*" )*).2$/#.+VVU*)/# *++*.$/ .$ *!/# ' !/>/# - 4*)./-$/$)"/# ' !/1$/4/*NY$)2$/#>*(+- 2$/#NZ$)# IWXJ)\E]$) IW>XJA* ''$)"/# '$")*)/*/# b(*' 0' $)*0-./-0/0- - 1 '.. 1 - ./ -$'.# .2$/#-*W[X!-*(E'**+)# VU]!-*(9WE8X'**+G$"0- VHA#$.> /*" /# -2$/#(0#- 0 . ' /$1$/4' !/1$/4*(+- /* >($"#/+- 1 )//# $)$)" *!/*#eA %$('+) %$%$ %$ $ $( )( $)C3B3<%#&"- 0-#e*(+' 3 .- 1 ')b(*' 0' *0)$) #.00)$/G$"0- WH>*+/$)" .. )/$''4$ )/$'*)!*-(/$*))$)/ -/$*)../# -/).*(/$# ./-0/0- .A $/$*)' ' /-*) ).$/42.*. -1 $)/# .0./-/ $)$)"+*& /% )//*/# *!/*-E $)$)".$/ G$"0- WH)2.- !$) ./2**0)+#*.+#/ $*).+ -.00)$/>+- .0('4 *-$"$)/$)"!-*(/# -4./''$5/$*)0!! -A# +*.$/$*).*!/# /2*+#*.+#/ $*).- $ )/$'$)*/# .00)$/.>)(+/2*)$*)$)$)".$/ .$)#/#/- *((*)'4*. -1 $)*/# - . G$"0- WHA# !$-./+#*.+#/ .$/ >&)*2)+- 1$*0.'4./# ..$/ >$.% )//*/# )$*/$)($ ) -$*. (*$ /$ .*!bA# *0)+#*.+#/ $)/ -/.2$/#- .$0 . )'*.$)"/# ..$/ G#-WZY> #-WZ[>-"XU[H.2 ''./# )$*/$)($ -$*. #4-*34'"-*0+*!b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eEbEY./-0/0- >/# /$1 .$/ . "( )/GW]UEW^WH*+/. /# B$)C*)!*-(/$*)!*-.00)$/>)($3 B$)CEB*0/C*)!*-(/$*)GZUa*0+)4 #H!*- .00)$/G$"0- WHA# /2**)!*-( -.$!! -./-0/0-''4$)/# *-$ )//$*)*!/# *). -1 - .$0 .#-W]X)'4W]Y>2#$#- $.+' !-*(/# B$)CG'*. -/*/# . *)+#*.+#/ H/*/# B*0/CG24!-*(/# . *)+#*.+#/ H*)!*-(/$*).4NWAUEXA]G8EHA)/ - ./$)"'4>/# . *)+#*.+#/ !-*(*/#.00)$/.*0+4/# B) 2$.$/ CG .+$/ +- . ) *!/# B*0/CE*)!*-( - $).00)$/H>)- )#*- 4 -WWX>#-WWZ) $.WZVG$"0- W>!#+) 'HA# B$)CE *)!*-( -*!.00)$/.)!0-/# -*)/-$0/ .#4-*" )*)./*/# . *)+#*.+#/ $*)1$/# ($)E#$)#-W]X)'4W]Y/*(.GNWAZEWA\H>2#$#- )*/+-*1$ $)/# .00)$/B*0/CE *)!*-( -./# /2*- .$0 .- (*- $./)/!-*(/# . *)+#*.+#/ GXA\)ZA\- .+ /$1 '4H G$"0- W>*(+- #)!#HA ] Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t $ $%".'%" $)$ %$'%$ ) %$( )( /-0/0-'#-/ -$5/$*)*!#eEb-4./'"-*2)$).0'+#/ E*)/$)$)"*)$/$*)- 1 '. ' /-*) ).$/4+ &.//# /2*)$*)$)$)".$/ .G$"0- XH/#/*)*/!$//# .#+ *!.0'+#/ $*).*!/ )*. -1 /*$)$)1-$*0. ./-0/0- .A)./ >/#$. ).$/4$."**(/#/* "'4 -*'+- . )/$)*0-+0-$!$/$*))-4*E+-*/ /)/0!! -.A.- .0'//2*"'4 -*'(*' 0' .2 - - !$) $)/*/# .)B) 2$C.$/ .*! #.00)$/>)2 - ! -- /*/#$../-0/0- ./# #eE bE*'*(+' 3A $/# -*!/# /2*"'4 -*'(*' 0' .*1 -'+.2$/#/# 3+ / '*/$*)*!/# .0./-/ EX"'4 -' #4 (*$ /4 /2 )/# /2*)$*)- *")$/$*).$/ .G$"0- XHA# $)/ -/$*). /2 )/# /2*"'4 -*'(*' 0' .)/# )54( - .$($'-/*/# +#*.+#/ $*).!-*( /# #eEbEY./-0/0- >' $/' .. 3/ ).$1 /#)/# +#*.+#/ $*)..- !' / 4 "- *!./-0/0-' " ) -4)!' 3$$/4$)/# $)$)"*)!*-(/$*)*!"'4 -*'A'/#*0"#$)/#$. ./-0/0- /# /$1 .$/ . "( )/*+/.!0''4/# B*0/C*)!*-(/$*)>/# "'4 -*'//# ) 2$.$/ $. !0-/# -24!-*(/# /'4/$4./ $) G*(+- /*/# +#*.+#/ $)/# #eEbEY *(+' 3H )'$)"$//*!*-($- /)2/ -E( $/ +*'-$)/ -/$*).2$/#- .$0 .!-*(/# /$1 .$/ . "( )/G -W]W>#-W]X)'4W]YHG$"0- XHA# 2/ -E( $/ #4-*" )*)) /2*-& -$"$)"/# /2*"'4 -*'(*' 0' .$.'.**). -1 $)/# #eEbEY./-0/0- A %#&') +! $) $".( (%C # ./ 4E.// &$) /$+-*+ -/$ .*!#e2 - ( .0- )*(+- 2$/#/#*. *!.*(/$ # G' WHA# /2*$.*54( ..#- .$($'-1'0 .!*-EX.0./-/ )-/ *)./)/A *2 1 ->#e#.XE!*''*2 -!*-bGXZ;H/#)# GVUU;H># ) - .0'/$)"$) )++-*3$(/ '4XE!*'$)- . /'4/$ !!$$ )4G/DH2$/#- "-./*bA# ./ 4E.// &$) /$.!*-# $)/#$../04- *(+-' /*/#*. +- 1$*0.'4- +*-/ !*-# +0-$!$ !-*( -$)/$..0 IW]J>.#*2$)"/*!NVUU.EV)*!UAU\()UAV[(!*-b)EX> - .+ /$1 '4A ^ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t ('(%$('+()'*)*'")*'(, )%)'( - 2 - +*-//# !$-./ /$' ./-0/0-')&$) /$#-/ -$5/$*)*!#0().+ -(E.+ $!$A 4!0.$)"E/ -($)'!!$)$/4/")/-0)/$)"/# E/ -($)'+*'4+-*'$) - "$*)/#/#$) -.*-- / +-*/ $)!*'$)"..#*2)$)/#$.2*-&>)$)- )/- +*-/IYVJ>2 #1 */$) #$"#' 1 '.*! - *($))/.*'0' #eA0-2*-&.$")$!$)/'4 3/ ).- )/./-0/0-'./04*)-/ IW^J>2# - 1 -4'*2- *($))/ 3+- ..$*)' 1 '.).*'0$'$/4- ,0$- *(+' 3/$*)2$/#)/$1 ) !*--4./''$./$*)A-*..E.+ $ .# / -*E/ /-( -.$)1-$*0../*$#$*( /-$ .#1 ) - +*-/ /2 ) ).00)$/.+- 1$*0.'4IW^>YWJA 1 -/# ' ..>"$1 )/# +-*3$($/4*! /# /$1 .$/ ' !//*/# .00)$/$)/ -! .>$/$.0)'$& '4/#/./0$ .*!$)/# . 0))/0-' # / -*E/ /-( -$!*-(.+-*1$ - '$./$.. ..( )/*!./-0/0- )/$1$/4A0-#$"#E - .*'0/$*)#e./-0/0- .*)!$-(#*(*E/ /-( -$!*-(/$*)).#*2./#/ ' /-*.//$ $)/ -/$*).)#-" *(+' ( )/-$/4++ -/*+'4.0./)/$'-*' $)/ /-( -$.. ('4>(*- .*/#)$)# >/*!$'$// /# *-- /*-$ )//$*)*!.00)$/.A0-./-0/0- .'.*- 1 ') !! /$1 '4'*. . ' /$1$/4' !/$)#.*(+- /**/# - .>+-/$0'-'4/# /-4+)*.*(' )54( .>)/# - !*- ' )!0-/# -.0++*-//*/# ./-/ "4*! .$")$)".+ $ .E.+ $!$ $)#$$/*-./#/!$''/# /-4+)*.*('. ' /$1$/4' !/2$/#*0/$)#$$/$)"/# #0() )54( .IW>XJA && $%$ %$'%$ ) %$( )( $ # $)$)"+*.$/$*).*!+#*.+#/ $)*0-#eEbEY*(+' 3(+/# /2*)$*)- *")$/$*) .$/ .$)#>2#$#- *0+$ 4"'4 -*'$)*0-#eEb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eEbEY./-0/0- ' -'4$)$/ ./#/+#*.+#/ $*)$.*0)//# ) 2$.$/ 2# )/# /$1 .$/ . "( )/*+/. /# !0''4B$)C*)!*-(/$*)G+- .0('4($($&$)"/ -)-4.// *! )54( +*$. !*-/'4.$.H)/# +-/$''4B*0/C*)!*-(/$*)G+- .0('4''*2$)"!'$++$)"*!/# # ($E /'$)/ -( $/ H>2#$' 2 $ )*/*. -1 +#*.+#/ $)$)"//# B*-$"$)'C$.$/ A#$.*)/-./.2$/#/# " / -)-4 *(+' 3>&)*2)/*#1 */#) 2)*-$"$)'$.$/ .*(+ / )/)+-/$''4*0+$ 4/2* +#*.+#/ (*$ /$ .>2$/#/# /$1 .$/ . "( )/*+/$)"/2**)!*-(/$*).IW\JA/$.)*/' - 2# /# -*E 3$./ ) *!/2*'/ -)/$1 $+*.$/$*).+*-/-4.*((*)/'4/$+-*+ -/4*-.+ $ .E .+ $!$! /0- IW\JA$) /#$.$.)*//# . $)#>$!! - )/( #)$.(!*-/# #0() )54( . ) )1$." 2# - /# *-$"$)'$.$/ (4)*/ +- . )/A VU Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d /#)& *-"$) --$" )*#'&!*-..$./) $)(...+ /-*( /-4)'4. .>)./!!/ /# $(*)$"#/*0- !*- ../*.4)#-*/-*)/*'' /$*)!$'$/$ .A ce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c THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t $) '$(),$$#.' ('%#")'%()) &'%&') ( $.- "-$)"/# 0)$,0 +*'4+-*'$) E/ -($)0.$)#>/# ($)*$$!! - ) . /2 ) #)# - (*./'4*)!$) /*/# +-*/ $).0-! >*)/-$0/$)"/*$!! - ) .$)/# $- .0-! ' /-*.//$+*/ )/$'.A# /$1 .$/ -#$/ /0- >'$")$)$)"(* )/'4/$- .$0 .> #*2 1 ->- #$"#'4*). -1 (*)"/# $.*54( .>$)+-/- !' / 4/# $-&$) /$+-( / -. $)" $)/# .( *- -*!(")$/0 A 1 -/# ' ..>.0/' $!! - ) .*1 ./)- 1$/$*).- )*/' > +-/$0'-'4/# (* -/ XE!*'#$"# -/'4/$ !!$$ )4!*-#/#)# A#$.(4 //-$0/' /*/# !! /*!($)*$.0./$/0/$*).+ -$+# -'/*/# /'4/$ )/- >*!/ )&)*2). BW).+# - C- .$0 .!*- )54( .&)*2)/*#1 $ )/$'/$1 .$/ .0/$!! - )/ )54(/$+-*+ -/$ . IYX>YYJA)0( -*!($)*$+*.$/$*).'$)$)"/# )$) ))$*/$)($ +*& /.*!/# bE $)$)".$/ G$"0- YH$)1*'1 +*$)/#-" #)" . /2 )#)# A*- 3(+' >4-V\X ).+XVV$)#- .0./$/0/ 4# ).)- .+ /$1 '4$)# G$"0- YHA# . BW) .+# - - .$0 .>2#$' )*/+ -/0-$)"/# ./-0/0-'$)/ "-$/4*!/# /$1 .$/ >*0'$)!'0 ) /# /'4/$ !!$$ )4*!/# )54( 4+*..$'4- (*1$)"#4-*" )*)2$/#bG4-V\X# .0./$/0/$*)H*-4'/ -$)"/# +)*). ,0 )/'4/# +-*/*)/$*).// G.+XVV.).0./$/0/$*)H*! ) -4/'4/$- .$0 .A **)'0 >$.*!*-(E.+ $!$$)#$$/*- 1 '*+( )/!*-##. )$(+' ( )/ !*-. 1 -' .../-/ "4!*-)*)E#*-(*)'*)/- +/$*)>/#*0"#.0!! -.!-*('&*!'$")E*0) ./-0/0-'$)!*-(/$*)!*-/# #0() $.*54( .A0-/#$"#'$"#/.0/' 4 /.$")$!$)/ $!! - ) .$)/# ./-0/0-')&$) /$! /0- . /2 )#)*/# - )54( .>) /# - !*- ) /* *).$ - !*-!0/0- .$")*!$)#$$/*-./-" /$)"/# .+ -(E.+ $!$$.*54( A VV Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t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icenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t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icenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t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icenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 "9)%"") %$$ $#$))) () (%'C()'*)*'( C3B3 C3B3< X#^ X+!2 $(*)$"#/*0- >UX $"&0E0+ --$"#/ UA^\[X VAZYV] W W []AWXFVA\WGVA]VFVA\WH WXAY^FWAVZGWAW\EWAVZH cVYWAU>-\VA]>c]VAX cVYXA[>-\VA^>c]VAV 8c:c^UAUR>9cVWWA^R 8c:c^UAUR>9cVWXAXR *A0)$,0 - !' /$*). \W>WW\GVU>WXUH XZ>XU]GZ>UUVH *(+' / ) ..GaH ^^AWG^[AXH ^YAVG^WAUH D< VVAYGWAXH \AUGWAXH ( -" GaH \A\GZVAYH V\AVG[\AVH 0))4 YAYGXA[H ZAXGZAXH *A/*(.$)- !$) ( )/GDDH ZXUVDVU]DZWX ZV^YDVWYDXVU -4./GaH VYAX V[A] !- GaH V\A[ WWAW E!/*-GDDHGWH W[DWUDW^ W]DXWDXU *)' )"/#GH UAUVY UAUV[ *))"' GRH VAZU VAZY 1*0- - "$*) ^\AU ^ZA^ ''*2 - "$*) ^^A\ ^^A\ '0 .$)-& /..#*2/# .//$./$.!*-/# #$"# ./- .*'0/$*).# ''.A DD )*/ +-*/ $)>/$1 E.$/ '$").)*/# -.*'1 )/(*' 0' .>- .+ /$1 '4 >-**/E( )E.,0- 1$/$*)A t sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 %#&"- ('$) 1 ' )"/#GH + "-*0+ .*'0/$*)-)" GH )$/ ''$( ).$*). VZ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 # VUUdX] UAWZdUAU^K GUAUWUHKK V^^dV\ (3G;(*' D($)D("H YUZdYV XYUdXV sc rip /G.EVH #e XZd] UAY[dUAV]K GUAUX\HKK WXYdWY bG;H EXG(H t ":).3())! $) &'%&') (C$2 Ac ce pte d Ma nu THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 '0'/$*).*!!*-EX2 - . $/# -*)K)..0(+/$*)*!) ,0'*) )/-/$*)*!E)E X$)/# - ($EX>*-KK)++-*3$(/ YaEX./# /$1 ' #4 $)/# - ($ .0./-/ *-$)"/*IYZJ>#*2 1 -/# '// -$.)*/*).$ - $)*/# - &$) /$)'4. .A V[ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t *'9)'*)*'"%+'+ ,1(*'")'%()) &%)$) "($(") + ).")%CAGH# .4(( /-$0)$/*!#eEbEY*(+' 3*(+-$. ..00)$/.)> #*(+*. *!/# E G"- )H)E*($).G4)HG"#HAGH# #e#*(*E/ /-( -$... (' !-*(/# $( - )/# $( -" ) -/ 4/2*E!*'-4./''*"-+#$.4(( /-4*+ -/*-G"- 4+') H'4$)"*)/# +') AGH0-! ' /-*.//$+*/ )/$'.*!#e'0'/ 4+-*"-(IY[J- 1 '/#- +-*($) )/'4#-" - "$*).@/# ) "/$1 '4E#-" E'**+G'0 H>+*.$/$1 '4E#-" XZUEX\W- "$*) G- H)($3E#-" VVUEVXU- "$*)G*-)" HA# "#.#*2.'0'/ ' /-*.//$+*/ )/$'.!*- .*(/$# G$V0]!HAGH0+ -+*.$/$*)*!/# #e. ' /$1$/4' !/$)'*. *)!*-(/$*)G4 ''*2&*) H2$/#/2*./-0/0- .*!.*(/$# ..#*2$)"/# '*. G'0 > $V0]!H)*+ )*)!*-(/$*).G*-)" >$V5),HA# '*. *)!*-(/$*)$)#$../$'$5 42/ -E( $/ #4-*" )*).G'0 .# '$) .>2/ -$)4 ''*2.+# - H>.$($'-/*/#/. )$) # G"- ).# '$) .>2/ -$)'0 .+# - .HAGH+ E!$''$)"- +- . )//$*)*!/# #e . ' /$1$/4' !/G4 ''*2H- 1 '.(0#- 0 1$/4/#) G(" )/>$V$XWH0 /*/# 0'&4# VU]>-*W[X)-"W[ZA# /-4+)*.*(' $)#$$/*-G*-)" H$.(* '' $)/* /# #./-0/0- . *)/# $)$)"+*.$/$*)*!/# )$) -$)"*!/# b*!/*-) $''0./-/ .. 1 -''.# . /2 )/# '$"))/# +-*/ $)A *':%(&) $ $( )( $C2GH !$) WE(+G*)/*0- /VAU<H!*-b) +#*.+#/ $*).$)/# #eEbEY*(+' 3AGH0+ -+*.$/$*)*!+#*.+#/ E*0) G. ($E/-).+- )/B*>$V"4+H).0'+#/ E*0)" G. ($E/-).+- )/B"*>$V"VH *)/*#eEbEY.00)$/G"- 4H(+./# /2**0)+#*.+#/ .$)#eG"- 4./$&.H/ /# ..$/ >)//# ) 2$.$/ A# +*.$/$*).*!/# *-$"$)')) 2$.$/ .$ )/$!$ !-*(1-$*0. ./-0/0- .- .#*2)AGH)/ -/$*).*!+#*.+#/ .//# .G#H)) 2$G!#H.$/ .A*/ /#///# ) 2$.$/ >/# '*. ./+#*.+#/ *34" )/*($.XAX24!-*(/# 4.WYY/#$*'"-*0+AGH 2**)!*-(/$*).*!/# /$1 .$/ . "( )/$)#eEbEY*(+' 3@)B$)C*)!*-(/$*)$) .00)$/G#H>)($3 B$)CG. ($E/-).+- )/HEB*0/CG"- 4H*)!*-(/$*).$).00)$/G!#HA *';".'%" $ $ $)C3B3%"%#&"-2GH #>,$!! - ) ).$/4$)/# /$1 .$/ *!*/#.00)$/.G*)/*0- /X<H2.(* '' ."'4 -*'A#(#*.+#/ )$*).!-*( /# #eEbEY./-0/0- G. ($E/-).+- )/H>)/2*EX(*' 0' .G*// /-).+- )/H *) *0)//# ..$/ GB!"*>!-*(-/EbEX*(+' 3>$W141H)/# */# -//# $ .$/ GB*>!-*( EbEX*(+' 3>$X(H- *1 -'$AGH)/ -/$*).*!/# /2* "'4 -*'(*' 0' .G*'H$).00)$/.G# H)G##H*!#eA/ -(*' 0' .- - +- . )/ .'0 .+# - .>)#4-*" )*)) /2*-&...# '$) .A*/ /# 2/ -E( $/ #4-*" )*)) /2*-&-$"$)"/# /2*"'4 -*'(*' 0' .$.#$"#'4*). -1 (*)"1-$*0.$)$)" *)!*-(/$*).*!"'4 -*'A *'<# $% (*() )*) %$(&' &'")%)) +( )(),$C$2GH ($)*$$!! - ) . /2 )#e)# - '0./ - $)/2*- "$*).*!/# /$1 .$/ @ *) % )//*/# )$) $)$)"+*& /)/# */# -//# )$*/$)($ $)$)"+*& /A .$0 ' '.- *'*0- '0 G.$H>- G$$H>"- )G0)#-" +*'-H*-"- 4G)*)E+*'-HAGH* -/ $!! - ) .$) ' /-*.//$+*/ )/$'.*!/# *!/*-+*& /)/'4/$+*& / /2 )#e )# - $-' A V\ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 *'9 sc rip nu Ma pte d ce Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t V] Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 *': pte d Ma nu THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 sc rip t Ac ce V^ Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 *'; Ac ce pte d Ma nu THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 sc rip t WU Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited Biochemical Journal Immediate Publication. Published on 26 Jan 2011 as manuscript BJ20101442 *'< sc rip nu Ma ce pte d Ac THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20101442 t WV Licenced copy. Copying is not permitted, except with prior permission and as allowed by law. © 2011 The Authors Journal compilation © 2011 Portland Press Limited
© Copyright 2024 Paperzz