2024-03-13RELRCSBRCSBRCSB2018-10-232018-11-072019-08-072024-03-13National Health and Medical Research Council (NHMRC, Australia)AustraliaAustralian Research Council (ARC)AustraliaThe structure of the Plasmodium falciparum 20S proteasome in complex with one PA28 activator.Hanssen EXie SCLeis AMetcalfe RDGillett DLTilley LGriffin MDWproteasome, protease, HYDROLASE, 11S subunit, hydrolyse activator, proteasome activator, complexXie SCMetcalfe RDHanssen EYang TGillett DLLeis APMorton CJKuiper MJParker MWSpillman NJWong WTsu CDick LRGriffin MDWTilley LThe structure of the PA28-20S proteasome complex from Plasmodium falciparum and implications for proteostasis.Nat MicrobiolUK419902000201931384003doi:10.1038/s41564-019-0524-42058-5276EMD-9258other EM volumeEMD-9257other EM volume6muxFULLOVERLAP20S proteasome/PA28 complex.20S proteasome/PA28 complex.0123456789101112131415Sample was a mixture of unbound 20S proteasome, 20S proteasome in complex with one PA28 cap, and 20S proteasome in complex with two PA28 caps.Plasmodium falciparum 3D70.23Plasmodium falciparum 20S proteasome11234567891011121314Plasmodium falciparum 3D711S activator of the Plasmodium falciparum proteasome.115Plasmodium falciparum 3D720S proteasome alpha-1 subunitPlasmodium falciparum 3D7isolate 3D70.0295316559999999962LEVOMVRPSQSMYDRHLTIFSPDGNLYQIEYAIKAVKNTNITSVGVKGENCAVIISQKKMATQYISQDKLLDYNNITNIYNITD
EIGCSMVGMPGDCLSMVYKARSEASEFLYSNGYNVNAETLCRNICDKIQVYTQHAYMRLHACSGMIIGIDENNKPELFKF
DPSGFCAGYRACVIGNKEQESISVLERLLEKRKKKIQQETIDEDIRNTTILAIEALQTILAFDLKASEIEVAIVSTKNRN
FTQISEKEIDNYLTYIAERDQ8IAR33.4.25.120S proteasome alpha-2 subunitPlasmodium falciparum 3D7isolate 3D70.0265563912LEVOMADGEYSFSLTTFSPTGKLVQIEYALNRVSSSSPALGIRAKNGVIIATEKKSPNELIEENSIFKIQQISEHIGIVYAGMP
GDFRVLLKRARKEAIRYSLQYGSEILVKELVKIIASIVQEFTQTGGVRPFGLSLLICGVDVYGYHLYQIDPSGCYFNWMA
TCVGKDYQNNMSFLEKRYNKDIEIEDAIHTAILTLKESYEGVLNEKNIEIGVAYDNKPFKILTQNEIKDYLIEIEC6KST33.4.25.120S proteasome alpha-3 subunitPlasmodium falciparum 3D7isolate 3D70.0279776642LEVOMARRYDSRTTTFSPEGRLYQVEYALEAINNASITIGLITKDGVILGADKVFISKLIDKANNYEKIYKIDKHIFCGVAGLN
ADANILINQSRLYAQRYLYNYNEVQPVSQLVVQICDIKQSYTQYGGLRPYGVSFLIGGYDTKDGYQLYHTDPSGNYSGWF
ATAIGTNNLTASSVLKQEWKNDMTLEEGLLLALKTLAKSTDTEIPKSEKIELAYLTNKDGEVYQKYLTEKEIEELIKLYT
QKYIKEQ8IDG33.4.25.120S proteasome alpha-4 subunitPlasmodium falciparum 3D7isolate 3D70.0272632849999999982LEVOMSYDRAITVFSPDGHLLQVEHALEAVKKGGCAVAIKSSNFAVLAVEKKNIPKLQNPKTTEKLIKLDEHNCLAFAGLNADA
RVLVNKTRLECQRYYLNMDEPAPVDYIAKYVAKVQQKFTHRGGVRPFGIATLIAGFKNNKEICIYQTEPSGIYAAWKAQA
IGKNAKIVQEFLEKNYQENMEQKDCIFLALKAIFEVVELSSKNVEVALLTEKDLTFIEEQEINSMVELIDQERTKNNEQN
EQ8IDG23.4.25.120S proteasome alpha-5 subunitPlasmodium falciparum 3D7isolate 3D70.0284173669999999952LEVOMFSTRSEYDRGVNTFSPEGRLFQVEYALGAIKLGSTAVGICVNDGVILASERRISSTLIEKDSVEKLLSIDDHIGCAMSG
LMADARTLIDYARVECNHYKFIYNENINIKSCVELISELALDFSNLSDSKRKKIMSRPFGVALLIGGVDKNGPCLWYTEP
SGTNTRFSAASIGSAQEGAELLLQENYKKDMTFEQAEILALTVLRQVMEDKLSTSNVEICAIKKSDQTFYKYNTDDISRI
IDVLPSPVYPTIDMTAQ8IBI33.4.25.120S proteasome alpha-6 subunitPlasmodium falciparum 3D7isolate 3D70.0288716969999999982LEVOMYRNLYDTDNIIYSPEGRLYQVEYASEAIKQGTCAVAIKSKDYVVVSGLKKCISKLSFPQEKIFKIDDYIGISMSGITSD
AKVLTKFMQNECLSHKFLYNENINIESLVRSVADKYQKNTQKSSKRAFGVGLMIAAYHNEPCIFETRPNGSYFEYDALSF
GARSHASKTYLEKNLHLFEECSLEELILHCLKALKCSLSSESELTISNTALAVVGKNHPWQEISSLQLEEYLSKVKMDAE
QEQVEENVQNEANEQ8IK903.4.25.120S proteasome alpha-7 subunitPlasmodium falciparum 3D7isolate 3D70.0293242949999999972LEVOMAGLSAGYDLSVSTFSPDGRLYQVEYIYKSINNNNTALCLECKDGIICCCINSNMDKNKMIKKNSYNRIYHVNNNIIITY
SGFDGDARNIIDRARSEANTYYYNFHTNIPLHILVNRISLYIHAYTLYWHMRPFAASIIISSFNEKDKGDIYCIEPNGAC
YKYSGIVIGKNKEMFKTEIEKKDYKDINVRDAIEDIYKFILTSDDHMNKNNLQNLVNFSWICKESSYEFQNIHEEILTPA
LNKAVEYIEKLNO773963.4.25.120S proteasome beta-1 subunitPlasmodium falciparum 3D7isolate 3D70.0291439362LEVOTTIIGIIYDNGVMLACDSRTSSGTFISNKCSRKINRINENLYVCRSGASAHSQKIIEIIKHYCVSMKNENRKKGRFHEGE
TIYDETTYDEEIDIDSINYLDYNNNNDNNLVTKNKYFYEDKFNDYNPLVENVAHITKKIIYTNNNFLSCALIFGGYDKIK
KQQLYAVNLNGSIIEKHDFAVSGSGSIYIQSYLQDKYKKFMTKKECFNLILNCVKYAMHNDNSSGGLIRIVNITKSFVEE
FTVVNTQMNFQYQ8I0U73.4.25.120S proteasome beta-2 subunitPlasmodium falciparum 3D7isolate 3D70.0251048849999999972LEVOTTICGLVCQNAVILGADTRATEGPIVADKNCSKLHYISKNIWCAGAGVAGDLEHTTLWLQHNVELHRLNTNTQPRVSMCV
SRLTQELFKYQGYKVCAIVLGGVDVNGPQLYGIHPHGSSCLLPFTALGSGSLNAMAVLEAKYRDNMTIEEGKNLVCEAIC
AGIFNDLGSGGNVDICVITKDSYQHIRPYKEPNMRLYHLPHPTIYPKGTTPILSEKIEYIKKFISVEDAQ8I6T33.4.25.120S proteasome beta-3 subunitPlasmodium falciparum 3D7isolate 3D70.0245331312LEVOMGSIYNYNGGCVLGMSGSNCVAIACDLRLGANTFTTVSTKFSKIFKMNNNVYVGLSGLATDIQTLYEILRYRVNLYEVRQ
DAEMDVECFANMLSSILYSNRFSPYFVNPIVVGFKLKHYVDEEGEKKVNYEPYLTAYDLIGAKCETRDFVVNGVTSEQLF
GMCESLYVKDQDENGLFETISQCLLSALDRDCISGWGAEVLVLTPEKIIKKKLKARMDQ8I2613.4.25.120S proteasome beta-4 subunitPlasmodium falciparum 3D7isolate 3D70.0228891052LEVOMDTLIGLRGNNFVVLAADTYSINSIIKLKNDDNTKFYDIHGNKCLLLGGSIGDRLQFGEFIRKNVHLYQYQNNTDMFVKS
FAFFTRKNLAYYLRRNPFEVNCLIAGYDKKDGYQLYWCDYLSNMDSVNKGAHGYGAYLVSAILDKYYHENLTVDEALDIF
KLCFEELKKRFLLTQINYELRIMYDNKVETQYVTVQ8IKC93.4.25.120S proteasome beta-5 subunitPlasmodium falciparum 3D7isolate 3D70.0236206462LEVOTTTLAFKFKDGIIVAVDSRASMGSFISSQNVEKIIEINKNILGTMAGGAADCLYWEKYLGKIIKIYELRNNEKISVRAAS
TILSNILYQYKGYGLCCGIILSGYDHTGFNMFYVDDSGKKVEGNLFSCGSGSTYAYSILDSAYDYNLNLDQAVELARNAI
YHATFRDGGSGGKVRVFHIHKNGYDKIIEGEDVFDLHYHYTNPEQKDQYVMQ8IJT13.4.25.120S proteasome beta-6 subunitPlasmodium falciparum 3D7isolate 3D70.0273012032LEVOMDLILYNDNLTEKKTEKENVIEHGRGFKRWYPYIDNGGTVIGLTGKDYVILAADTRLSLSYSIYTRFCPKISKLTDKCII
GSSGMQSDIKTLHSLLQKKIQLFVLEHSHYPDIHVIARLLCVILYSRRFFPYYAFNILAGVDENNKGVLYNYDSVGSYCE
ATHSCVGSGSQLILPILDNRVEQKNQLIKNTNFNLGDDINFVKDAITSATERDIYTGDKTLIYVIDKMGINVNTLDLKQDC0H4E83.4.25.120S proteasome beta-7 subunitPlasmodium falciparum 3D7isolate 3D70.0309098929999999972LEVOMTLGPVVTGTSVIAIKYKHGIMIAADRKASYGSYAKFQNVERIFKINNKTVMGFSGELADAQYLHELLTRKNINNLSEKK
RKEDMYTPQHYHSYVSRVFYVRKNRIDPLFNNIIIAGINSQKYDNNDDNVLLYTNKNNDDEQNEYKNNEEYKEIHKDDLY
IGFVDMHGTNFCDDYITTGYARYFALTLLRDHYKDNMTEEEARILINECLRILYFRDATSSNFIQIVKVTSKGVEYEEPY
ILPCVLNSADYVYPSTLLPPAGCMWQ7K6A93.4.25.1Proteasome activator PA28Plasmodium falciparum 3D7isolate 3D70.0331787737Escherichia coliLEVOMEAYLNKIDKIEPSDQKIKEEYNKFKYDITKQAIESLRERIPKRIIFFNNLVNVNSEPGSILNVNDLDGVSYKYKINKIE
ENVRKKHKGNNSSDFDEREKKKDSLDGHVKHFSNNEDSKLIIDDKVLYTHYVPSHKQIYLELEKIKTYASELIEIIGNIK
LWIQLNVPRIEDGNNFGVGIQEEAIQELARVEESAFNLYDAIVKYYMERAKISTKVLKYPNVSDYQEAVRELDEKEWIHI
KITIVDMRNNYIMLYDLLYKNWEKVVKPKNEDAHHRMTFQ8I374singleParticleparticle0.77.4100.0NaClSodium Chloride5.0MgCl2Magnesium Chloride1.0DTTDithiothreitol25.0Tris-HClTris hydrochlorideunspecifiedETHANE100277.15FEI VITROBOT MARK IVwait time 0sec
blot time 2sec
blot force -1. FEI TALOS ARCTICAFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN200100.02.71.03.0100000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGIF Bioquantum20GATAN K2 QUANTUM (4k x 4k)COUNTING383837101-401520010.032.01Images were gain corrected212749Relion autopick from 5 class averages resulting from 200o0 particle picked manuallyDe novo model created form 10,000 particle picked in Relion. No symmetry impliedC13.9FSC 0.143 CUT-OFFRELION3.057337MAXIMUM LIKELIHOODRELION2.1MAXIMUM LIKELIHOODRELION2.16RELION2.16DFKMPDBexperimental model6MUWPDBexperimental modelAB INITIO MODELREAL82.680000000000007emd_9259_additional_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
344
344344
0
00344344344450.63998450.63998450.6399890.090.090.0XYZ-0.0192917530.05853376-0.0000368686780.00229986781.311.311.31original unsharpened mapemd_9259_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
344
344344
0
00344344344450.63998450.63998450.6399890.090.090.0XYZ-0.0295045620.07129318-0.0000170616840.00328420821.311.311.31Plasmodium falciparum 20S proteasomeemd_9259_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)