MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins. The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins. EMBO J. (0261-4189) 2011 imex curation Dekker C., Roe SM., McCormack EA., Beuron F., Pearl LH., Willison KR. Only protein-protein interactions in vitro In vitro crosslink cross-linking study crosslink predetermined predetermined participant predetermined Dekker C., Roe SM., McCormack EA., Beuron F., Pearl LH., Willison KR. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins. The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins. EMBO J. (0261-4189) 2011 imex curation Dekker C., Roe SM., McCormack EA., Beuron F., Pearl LH., Willison KR. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Dekker C., Roe SM., McCormack EA., Beuron F., Pearl LH., Willison KR. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins. The crystal structure of yeast CCT reveals intrinsic asymmetry of eukaryotic cytosolic chaperonins. EMBO J. (0261-4189) 2011 imex curation Dekker C., Roe SM., McCormack EA., Beuron F., Pearl LH., Willison KR. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Dekker C., Roe SM., McCormack EA., Beuron F., Pearl LH., Willison KR. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation tcpd_yeast T-complex protein 1 subunit delta YDL143W CCT4 ANC2 TCP4 CCT-delta protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAKVPSNATFKNKEKPQEVRKANIIAARSVADAIRTSLGPKGMDKMIKTSRGEIIISNDGHTILKQMAILHPVARMLVEVSAAQDSEAGDGTTSVVILTGALLGAAERLLNKGIHPTIIADSFQSAAKRSVDILLEMCHKVSLSDREQLVRAASTSLSSKIVSQYSSFLAPLAVDSVLKISDENSKNVDLNDIRLVKKVGGTIDDTEMIDGVVLTQTAIKSAGGPTRKEKAKIGLIQFQISPPKPDTENNIIVNDYRQMDKILKEERAYLLNICKKIKKAKCNVLLIQKSILRDAVNDLALHFLSKLNIMVVKDIEREEIEFLSKGLGCKPIADIELFTEDRLGSADLVEEIDSDGSKIVRVTGIRNNNARPTVSVVIRGANNMIIDETERSLHDALCVIRCLVKERGLIAGGGAPEIEISRRLSKEARSMEGVQAFIWQEFASALEVIPTTLAENAGLNSIKVVTELRSKHENGELNDGISVRRSGTTNTYEEHILQPVLVSTSAITLASECVKSILRIDDIAFSR F01146D76F4E850C F01146D76F4E850C plp2_yeast Phosducin-like protein 2 Viral IAP-associated factor 1 homolog PLP2 VIAF1 YOR281C protein protein yeast Saccharomyces cerevisiae Baker's yeast MQNEPMFQVQVDESEDSEWNDILRAKGVIPERAPSPTAKLEEALEEAIAKQHENRLEDKDLSDLEELEDDEDEDFLEAYKIKRLNEIRKLQERSKFGEVFHINKPEYNKEVTLASQGKKYEGAQTNDNGEEDDGGVYVFVHLSLQSKLQSRILSHLFQSAACKFREIKFVEIPANRAIENYPESNCPTLIVYYRGEVIKNMITLLELGGNNSKMEDFEDFMVKVGAVAEGDNRLIMNRDDEESREERKLHYGEKKSIRSGIRGKFNVGIGGNDDGNINDDDDGFFD 414A696FFBDBD833 414A696FFBDBD833 tcpa_yeast T-complex protein 1 subunit alpha TCP1 CCT1 YDR212W YD8142.13 YD8142B.04 CCT-alpha protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQLFNNSRSDTLFLGGEKISGDDIRNQNVLATMAVANVVKSSLGPVGLDKMLVDDIGDFTVTNDGATILSLLDVQHPAGKILVELAQQQDREIGDGTTSVVIIASELLKRANELVKNKIHPTTIITGFRVALREAIRFINEVLSTSVDTLGKETLINIAKTSMSSKIIGADSDFFSNMVVDALLAVKTQNSKGEIKYPVKAVNVLKAHGKSATESLLVPGYALNCTVASQAMPKRIAGGNVKIACLDLNLQKARMAMGVQINIDDPEQLEQIRKREAGIVLERVKKIIDAGAQVVLTTKGIDDLCLKEFVEAKIMGVRRCKKEDLRRIARATGATLVSSMSNLEGEETFESSYLGLCDEVVQAKFSDDECILIKGTSKHSSSSIILRGANDYSLDEMERSLHDSLSVVKRTLESGNVVPGGGCVEAALNIYLDNFATTVGSREQLAIAEFAAALLIIPKTLAVNAAKDSSELVAKLRSYHAASQMAKPEDVKRRSYRNYGLDLIRGKIVDEIHAGVLEPTISKVKSLKSALEACVAILRIDTMITVDPEPPKEDPHDH 02C625D329359E61 02C625D329359E61 tcpb_yeast T-complex protein 1 subunit beta YIL142W CCT2 BIN3 TCP2 CCT-beta protein protein yeast Saccharomyces cerevisiae Baker's yeast MSVQIFGDQVTEERAENARLSAFVGAIAVGDLVKSTLGPKGMDKLLQSASSNTCMVTNDGATILKSIPLDNPAAKVLVNISKVQDDEVGDGTTSVTVLSAELLREAEKLIDQSKIHPQTIIEGYRLASAAALDALTKAAVDNSHDKTMFREDLIHIAKTTLSSKILSQDKDHFAELATNAILRLKGSTNLEHIQIIKILGGKLSDSFLDEGFILAKKFGNNQPKRIENAKILIANTTLDTDKVKIFGTKFKVDSTAKLAQLEKAEREKMKNKIAKISKFGINTFINRQLIYDYPEQLFTDLGINSIEHADFEGVERLALVTGGEVVSTFDEPSKCKLGECDVIEEIMLGEQPFLKFSGCKAGEACTIVLRGATDQTLDEAERSLHDALSVLSQTTKETRTVLGGGCAEMVMSKAVDTEAQNIDGKKSLAVEAFARALRQLPTILADNAGFDSSELVSKLRSSIYNGISTSGLDLNNGTIADMRQLGIVESYKLKRAVVSSASEAAEVLLRVDNIIRARPRTANRQHM C5DB2BC3660EC7E6 C5DB2BC3660EC7E6 tcpe_yeast T-complex protein 1 subunit epsilon CCT5 TCP5 YJR064W J1752 CCT-epsilon protein protein yeast Saccharomyces cerevisiae Baker's yeast MAARPQQPPMEMPDLSNAIVAQDEMGRPFIIVKDQGNKKRQHGLEAKKSHILAARSVASIIKTSLGPRGLDKILISPDGEITITNDGATILSQMELDNEIAKLLVQLSKSQDDEIGDGTTGVVVLASALLDQALELIQKGIHPIKIANGFDEAAKLAISKLEETCDDISASNDELFRDFLLRAAKTSLGSKIVSKDHDRFAEMAVEAVINVMDKDRKDVDFDLIKMQGRVGGSISDSKLINGVILDKDFSHPQMPKCVLPKEGSDGVKLAILTCPFEPPKPKTKHKLDISSVEEYQKLQTYEQDKFKEMIDDVKKAGADVVICQWGFDDEANHLLLQNDLPAVRWVGGQELEHIAISTNGRIVPRFQDLSKDKLGTCSRIYEQEFGTTKDRMLIIEQSKETKTVTCFVRGSNKMIVDEAERALHDSLCVVRNLVKDSRVVYGGGAAEVTMSLAVSEEADKQRGIDQYAFRGFAQALDTIPMTLAENSGLDPIGTLSTLKSKQLKEKISNIGVDCLGYGSNDMKELFVVDPFIGKKQQILLATQLCRMILKIDNVIISGKDEY 8CDAF88C6403655F 8CDAF88C6403655F tcph_yeast T-complex protein 1 subunit eta CCT7 YJL111W J0804 CCT-eta protein protein yeast Saccharomyces cerevisiae Baker's yeast MNFGSQTPTIVVLKEGTDASQGKGQIISNINACVAVQEALKPTLGPLGSDILIVTSNQKTTISNDGATILKLLDVVHPAAKTLVDISRAQDAEVGDGTTSVTILAGELMKEAKPFLEEGISSHLIMKGYRKAVSLAVEKINELAVDITSEKSSGRELLERCARTAMSSKLIHNNADFFVKMCVDAVLSLDRNDLDDKLIGIKKIPGGAMEESLFINGVAFKKTFSYAGFEQQPKKFNNPKILSLNVELELKAEKDNAEVRVEHVEDYQAIVDAEWQLIFEKLRQVEETGANIVLSKLPIGDLATQFFADRNIFCAGRVSADDMNRVIQAVGGSIQSTTSDIKPEHLGTCALFEEMQIGSERYNLFQGCPQAKTCTLLLRGGAEQVIAEVERSLHDAIMIVKRALQNKLIVAGGGATEMEVSKCLRDYSKTIAGKQQMIINAFAKALEVIPRQLCENAGFDAIEILNKLRLAHSKGEKWYGVVFETENIGDNFAKFVWEPALVKINALNSATEATNLILSVDETITNKGSESANAGMMPPQGAGRGRGMPM A428FEC2ACD54B4B A428FEC2ACD54B4B act_yeast Actin ACT1 ABY1 END7 YFL039C protein protein yeast Saccharomyces cerevisiae Baker's yeast MDSEVAALVIDNGSGMCKAGFAGDDAPRAVFPSIVGRPRHQGIMVGMGQKDSYVGDEAQSKRGILTLRYPIEHGIVTNWDDMEKIWHHTFYNELRVAPEEHPVLLTEAPMNPKSNREKMTQIMFETFNVPAFYVSIQAVLSLYSSGRTTGIVLDSGDGVTHVVPIYAGFSLPHAILRIDLAGRDLTDYLMKILSERGYSFSTTAEREIVRDIKEKLCYVALDFEQEMQTAAQSSSIEKSYELPDGQVITIGNERFRAPEALFHPSVLGLESAGIDQTTYNSIMKCDVDVRKELYGNIVMSGGTTMFPGIAERMQKEITALAPSSMKVKIIAPPERKYSVWIGGSILASLTTFQQMWISKQEYDESGPSIVHHKCF 87AC19B0B0BC9E71 87AC19B0B0BC9E71 acts_rabit Actin, alpha skeletal muscle ACTA1 ACTA Alpha-actin-1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MCDEDETTALVCDNGSGLVKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGDEAQSKRGILTLKYPIEHGIITNWDDMEKIWHHTFYNELRVAPEEHPTLLTEAPLNPKANREKMTQIMFETFNVPAMYVAIQAVLSLYASGRTTGIVLDSGDGVTHNVPIYEGYALPHAIMRLDLAGRDLTDYLMKILTERGYSFVTTAEREIVRDIKEKLCYVALDFENEMATAASSSSLEKSYELPDGQVITIGNERFRCPETLFQPSFIGMESAGIHETTYNSIMKCDIDIRKDLYANNVMSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWITKQEYDEAGPSIVHRKCF DIP protein P68135 original sequence version: 41 DIP protein P68135 original sequence version: 27 DIP protein P68135 original sequence version: 28 DIP protein P68135 original sequence version: 96 DIP protein P68135 original sequence version: 29 DIP protein P68135 original sequence version: 89 DIP protein P68135 original sequence version: 39 DIP protein P68135 original sequence version: 42 DIP protein P68135 original sequence version: 59 DIP protein P68135 original sequence version: 55 DIP protein P68135 original sequence version: 60 DIP protein P68135 original sequence version: 65 DIP protein P68135 original sequence version: 67 DIP protein P68135 original sequence version: 63 DIP protein P68135 original sequence version: 68 DIP protein P68135 original sequence version: 79 DIP protein P68135 original sequence version: 88 DIP protein P68135 original sequence version: 84 DIP protein P68135 original sequence version: 101 DIP protein P68135 original sequence version: 94 DIP protein P68135 original sequence version: 95 DIP protein P68135 original sequence version: 97 DIP protein P68135 original sequence version: 100 DIP protein P68135 original sequence version: 102 DIP protein P68135 original sequence version: 83 DF2A3A046346A179 tcpg_yeast T-complex protein 1 subunit gamma CCT3 BIN2 TCP3 YJL014W J1336 CCT-gamma protein protein yeast Saccharomyces cerevisiae Baker's yeast MQAPVVFMNASQERTTGRQAQISNITAAKAVADVIRTCLGPKAMLKMLLDPMGGLVLTNDGHAILREIDVAHPAAKSMLELSRTQDEEVGDGTTTVIILAGEILAQCAPYLIEKNIHPVIIIQALKKALTDALEVIKQVSKPVDVENDAAMKKLIQASIGTKYVIHWSEKMCELALDAVKTVRKDLGQTVEGEPNFEIDIKRYVRVEKIPGGDVLDSRVLKGVLLNKDVVHPKMSRHIENPRVVLLDCPLEYKKGESQTNIEIEKEEDWNRILQIEEEQVQLMCEQILAVRPTLVITEKGVSDLAQHYLLKGGCSVLRRVKKSDNNRIARVTGATIVNRVEDLKESDVGTNCGLFKVEMIGDEYFSFLDNCKEPKACTIMLRGGSKDILNEIDRNLQDAMAVARNVMLSPSLSPGGGATEMAVSVKLAEKAKQLEGIQQWPYQAVADAMECIPRTLIQNAGGDPIRLLSQLRAKHAQGNFTTGIDGDKGKIVDMVSYGIWEPEVIKQQSVKTAIESACLLLRVDDIVSGVRKQE 5C5AFF3D67D9A3B6 5C5AFF3D67D9A3B6 tcpq_yeast T-complex protein 1 subunit theta CCT8 YJL008C J1374 CCT-theta protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLRLPQNPNAGLFKQGYNSYSNADGQIIKSIAAIRELHQMCLTSMGPCGRNKIIVNHLGKIIITNDAATMLRELDIVHPAVKVLVMATEQQKIDMGDGTNLVMILAGELLNVSEKLISMGLSAVEIIQGYNMARKFTLKELDEMVVGEITDKNDKNELLKMIKPVISSKKYGSEDILSELVSEAVSHVLPVAQQAGEIPYFNVDSIRVVKIMGGSLSNSTVIKGMVFNREPEGHVKSLSEDKKHKVAVFTCPLDIANTETKGTVLLHNAQEMLDFSKGEEKQIDAMMKEIADMGVECIVAGAGVGELALHYLNRYGILVLKVPSKFELRRLCRVCGATPLPRLGAPTPEELGLVETVKTMEIGGDRVTVFKQEQGEISRTSTIILRGATQNNLDDIERAIDDGVAAVKGLMKPSGGKLLPGAGATEIELISRITKYGERTPGLLQLAIKQFAVAFEVVPRTLAETAGLDVNEVLPNLYAAHNVTEPGAVKTDHLYKGVDIDGESDEGVKDIREENIYDMLATKKFAINVATEAATTVLSIDQIIMAKKAGGPRAPQGPRPGNWDQED 90817CA3151A52FD 90817CA3151A52FD tcpz_yeast T-complex protein 1 subunit zeta CCT6 TCP20 TCP6 YDR188W YD9395.21 CCT-zeta protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLQLLNPKAESLRRDAALKVNVTSAEGLQSVLETNLGPKGTLKMLVDGAGNIKLTKDGKVLLTEMQIQSPTAVLIARAAAAQDEITGDGTTTVVCLVGELLRQAHRFIQEGVHPRIITDGFEIARKESMKFLDEFKISKTNLSNDREFLLQVARSSLLTKVDADLTEVLTPIVTDAVLSVYDAQADNLDLHMVEIMQMQHLSPKDTTFIKGLVLDHGGRHPDMPTRVKNAYVLILNVSLEYEKTEVNSGFFYSSADQRDKLAASERKFVDAKLKKIIDLKNEVCGMDPDKGFVIINQKGIDPMSLDVFAKHNILALRRAKRRNMERLQLVTGGEAQNSVEDLSPQILGFSGLVYQETIGEEKFTYVTENTDPKSCTILIKGSTHYALAQTKDAVRDGLRAVANVLKDKNIIPGAGAFYIALSRYLRSANMNKLGAKGKTKTGIEAFAEALLVIPKTLVKNSGFDPLDVLAMVEDELDDAQDSDETRYVGVDLNIGDSCDPTIEGIWDSYRVLRNAITGATGIASNLLLCDELLRAGRSTLKETPQ 7D2A2AB4526D3DB4 7D2A2AB4526D3DB4 cct2-cct3 2 CCT3 12 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT2 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT8 13 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT6 14 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT7 9 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT4 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT5 8 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified physical association physical association homomint mint The CCT aactin complex was established in the presence of PLP2 that acts as a cofactor and stabilises CCT actin interaction (Stirling et al, 2007; McCormack et al, 2009; Supplementary Figure S1). The ternary complex of CCT Actin PLP2 was crystallised in the presence of ATP and beryllium fluoride. A 3.8-A° data set was collected at the Swiss Light Source; data statistics are shown in Supplementary Table I. f1a f1b st1 sf5 sf6 3.8 A R = 0.3, Rwork = 0.3, Rfree = 0.34 acta1-cct7 1 CCT1 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified CCT8 13 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Alpha-actin 11 weight by staining molecular weight estimation by staining weight by staining predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified CCT7 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified physical association physical association f5c mint Since the ternary complex of crystallised CCT Actin PLP2 was formed from rabbit a-actin, we repeated the crosslinking analysis with this complex (Figure 5C) and obtained similar results: crosslinks of a-actin to Cct1, Cct7 and Cct8. This crosslinking confirms that the rabbit actin binds CCT in a similar manner to yeast actin, namely near the apical domains of Cct1 and Cct7, which is compatible with much of the density we attribute to actin. act1-cct5 1 CCT8 13 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified CCT5 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified CCT7 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified CCT1 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Alpha 10 weight by staining molecular weight estimation by staining weight by staining predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified physical association physical association f5b homomint mint native yeast actin (ACT1), labelled with AlexaFluor488 on cysteine 374, was derivatised with a bifunctional crosslinker, succinimidyl-ester diazirine (sulfo- SDA), and CCT ACT1 PLP2 complexes were assembled by EDTA-mediated unfolding of actin in the presence of CCT and PLP2 (McCormack et al, 2009) and crosslinked using UV irradiation (Figure 5A). Samples were probed with all eight anti-CCT subunit antibodies as previously described (McCormack et al, 2009). Crosslinked species compatible with a crosslink formed between one chain of the respective CCT subunit and one yeast actin polypeptide were observed for Cct1, Cct5, Cct7 and Cct8 (Figure 5B). plp2-cct4 1 PLP2 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified CCT1 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified CCT8 13 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified CCT4 4 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified physical association physical association f6a homomint mint We performed further crosslinking experiments using sulfo-SDA PLP2 (Figure 6A) and three PLP2 deletion mutants (Figure 6B) which mapped interactions between PLP2 and Cct1, Cct4 and Cct8 (Figure 6C), identifying also a potential Cct4 Cct8 PLP2 complex (species 4). The PLP2 interaction with Cct1 and Cct8, across the ring equator, is consistent with previous EM models of CCT phosducin-like protein interactions (Martin-Benito et al, 2004). acta1-cct2 3 CCT2 7 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT6 14 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT4 4 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Alpha-actin 11 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT5 8 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT7 9 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified PLP2 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT1 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT3 12 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CCT8 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified physical association physical association sf1b mint