2022-08-24RELRCSBRCSBRCSB2021-10-152022-07-062022-07-062022-08-24National Institutes of Health/National Cancer Institute (NIH/NCI)R01CA221289United StatesNational Institutes of Health/National Cancer Institute (NIH/NCI)F31CA224804United StatesHuman Trio residues 1284-1959 in complex with Rac1Chen C-LRavala SKBandekar SJCash JTesmer JJGBandekar SJChen CLRavala SKCash JNAvramova LVZhalnina MVGutkind JSLi STesmer JJGStructural/functional studies of Trio provide insights into its configuration and show that conserved linker elements enhance its activity for Rac1.J.Biol.Chem.US298102209102209202235779635doi:10.1016/j.jbc.2022.1022091083-351X0071JBCHA3EMD-25153associated EM volumeHuman Trio residues 1284-1959 in complex with Rac17sj4FULLOVERLAPHuman Trio residues 1284-1959 in complex with Rac1Human Trio residues 1284-1959 in complex with Rac1012Homo sapiensEscherichia coliRosetta (DE3) pLysSTriple functional domain proteinHomo sapiens0.0755929691Escherichia coliLEVOSNAEEKRKSARRKEFIMAELIQTEKAYVRDLRECMDTYLWEMTSGVEEIPPGIVNKELIIFGNMQEIYEFHNNIFLKELE
KYEQLPEDVGHCFVTWADKFQMYVTYCKNKPDSTQLILEHAGSYFDEIQQRHGLANSISSYLIKPVQRITKYQLLLKELL
TCCEEGKGEIKDGLEVMLSVPKRANDAMHLSMLEGFDENIESQGELILQESFQVWDPKTLIRKGRERHLFLFEMSLVFSK
EVKDSSGRSKYLYKSKLFTSELGVTEHVEGDPCKFALWVGRTPTSDNKIVLKASSIENKQDWIKHIREVIQERTIHLKGA
LKEPIHIPKTAPATRQKGRRDGEDLDSQGDGSSQPDTISIASRTSQNTLDSDKLSGGCELTVVIHDFTACNSNELTIRRG
QTVEVLERPHDKPDWCLVRTTDRSPAAEGLVPCGSLCIAHSRSSMEMEGIFNHKDSLSVSSNDASPPASVASLQPHMIGA
QSSPGPKRPGNTLRKWLTSPVRRLSSGKADGHVKKLAHKHKKSREVRKSADAGSQKDSDDSAATPQDETVEERGRNEGLS
SGTLSKSSSSGMQSCGEEEGEEGADAVPLPPPMAIQQHSLLQPDSQDDKASSRLLVRPTSSETPSAAELVSAIEELVKSK
MALEDRPSSLLVDQGDSSSPSFNPSDNSLLSSSSPIDEM2.7.11.1Ras-related C3 botulinum toxin substrate 1Homo sapiens0.0218114531Escherichia coliLEVOGEFMQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDV
FLICFSLVSPASFENVRAKWYPEVRHHCPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLEC
SALTQRGLKTVFDEAIRAVLCPPPVKKRKRKCLLLwater1.8015e-0524HOHsingleParticleparticle0.258.020.0C8H18N2O4S(4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)200.0NaClSodium Chloride2.0C4H10O2S2dithiothreitolUltrAuFoil R1.2/1.3GOLD300GOLDHOLEYGLOW DISCHARGEETHANE100277FEI VITROBOT MARK IVBlot force 10. FEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN300100.02.71.03.081000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGEN0.01GATAN K3 BIOQUANTUM (6k x 4k)SUPER-RESOLUTION11520818428173.1255.03514 movies were initially collected. After motion correction and CTF estimation, 697 micrographs were rejected due to poor CTF fitting.1cryoSPARC3.3.1Ab-initio model generated from cryoSPARCC1BACK PROJECTION2.86FSC 0.143 CUT-OFFcryoSPARC3.3.1922202COMMON LINEcryoSPARC3.3.1COMMON LINEcryoSPARC3.3.12cryoSPARC3.3.12NZ8A2NZ8BRIGID BODY FITThe initial model was generated from the pdb structure (2NZ8) using the SWISS-MODEL server and rigid-body fitted using Chimera. Several runs of structure refinement were done using the coot and phenix real-space refinement.correlation coefficientREAL