IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Insights into Antiparallel Microtubule Crosslinking by PRC1, a Conserved Nonmotor Microtubule Binding Protein. Insights into Antiparallel Microtubule Crosslinking by PRC1, a Conserved Nonmotor Microtubule Binding Protein. Cell (0092-8674) 2010 imex curation Subramanian R., Wilson-Kubalek EM., Arthur CP., Bick MJ., Campbell EA., Darst SA., Milligan RA., Kapoor TM. Initial fluorescence intensity analysis to determine the oligomerization state of the GFP tagged PRC1 constructs was performed by immobilizing GFP-tagged protein nonspecifically to a glass coverslip. After washing away unbound protein, motility buffer was added to the chamber and images were acquired. Fluorescent spots were selected by visual inspection and the average intensity of the first five frames was used to compute the initial intensity after background subtraction. kapoor@mail.rockefeller.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Subramanian R., Wilson-Kubalek EM., Arthur CP., Bick MJ., Campbell EA., Darst SA., Milligan RA., Kapoor TM. Initial fluorescence intensity analysis to determine the oligomerization state of the GFP tagged PRC1 constructs was performed by immobilizing GFP-tagged protein nonspecifically to a glass coverslip. After washing away unbound protein, motility buffer was added to the chamber and images were acquired. Fluorescent spots were selected by visual inspection and the average intensity of the first five frames was used to compute the initial intensity after background subtraction. kapoor@mail.rockefeller.edu Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation Insights into Antiparallel Microtubule Crosslinking by PRC1, a Conserved Nonmotor Microtubule Binding Protein. Insights into Antiparallel Microtubule Crosslinking by PRC1, a Conserved Nonmotor Microtubule Binding Protein. Cell (0092-8674) 2010 imex curation Subramanian R., Wilson-Kubalek EM., Arthur CP., Bick MJ., Campbell EA., Darst SA., Milligan RA., Kapoor TM. Initial fluorescence intensity analysis to determine the oligomerization state of the GFP tagged PRC1 constructs was performed by immobilizing GFP-tagged protein nonspecifically to a glass coverslip. After washing away unbound protein, motility buffer was added to the chamber and images were acquired. Fluorescent spots were selected by visual inspection and the average intensity of the first five frames was used to compute the initial intensity after background subtraction. kapoor@mail.rockefeller.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Subramanian R., Wilson-Kubalek EM., Arthur CP., Bick MJ., Campbell EA., Darst SA., Milligan RA., Kapoor TM. Initial fluorescence intensity analysis to determine the oligomerization state of the GFP tagged PRC1 constructs was performed by immobilizing GFP-tagged protein nonspecifically to a glass coverslip. After washing away unbound protein, motility buffer was added to the chamber and images were acquired. Fluorescent spots were selected by visual inspection and the average intensity of the first five frames was used to compute the initial intensity after background subtraction. kapoor@mail.rockefeller.edu Accepted 2010-OCT-3 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation o43663-1 Protein regulator of cytokinesis 1 PRC1 protein protein human Homo sapiens Human MRRSEVLAEESIVCLQKALNHLREIWELIGIPEDQRLQRTEVVKKHIKELLDMMIAEEESLKERLIKSISVCQKELNTLCSELHVEPFQEEGETTILQLEKDLRTQVELMRKQKKERKQELKLLQEQDQELCEILCMPHYDIDSASVPSLEELNQFRQHVTTLRETKASRREEFVSIKRQIILCMEALDHTPDTSFERDVVCEDEDAFCLSLENIATLQKLLRQLEMQKSQNEAVCEGLRTQIRELWDRLQIPEEEREAVATIMSGSKAKVRKALQLEVDRLEELKMQNMKKVIEAIRVELVQYWDQCFYSQEQRQAFAPFCAEDYTESLLQLHDAEIVRLKNYYEVHKELFEGVQKWEETWRLFLEFERKASDPNRFTNRGGNLLKEEKQRAKLQKMLPKLEEELKARIELWEQEHSKAFMVNGQKFMEYVAEQWEMHRLEKERAKQERQLKNKKQTETEMLYGSAPRTPSKRRGLAPNTPGKARKLNTTTMSNATANSSIRPIFGGTVYHSPVSRLPPSGSKPVAASTCSGKKTPRTGRHGANKENLELNGSILSGGYPGSAPLQRNFSINSVASTYSEFAKDPSLSDSSTVGLQRELSKASKSDATSGILNSTNIQS DIP protein O43663-1 original sequence version: 118 DIP protein O43663 original sequence version: 75 28393C5B1B3662F3 prc1-1 2 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain n-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction For visualizing PRC1 molecules by fluorescence microscopy we expressed and purified recombinant GFP-tagged full-length PRC1 in bacteria. Fig. 1A prc1-2 1 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain n-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction HPLC Size Exclusion Chromatography/ Laser Light Scattering analysis of PRC1-NSΔC (aa.1-486). Supp. Fig. 3