title: Expression, Purification, and Characterization of Interleukin-11 Orthologues.
abstract: Interleukin-11 (IL-11) is a multifunctional cytokine implicated in several normal and pathological processes. The decoding of IL-11 function and development of IL-11-targeted drugs dictate the use of laboratory animals and need of the better understanding of species specificity of IL-11 signaling. Here, we present a method for the recombinant interleukin-11 (rIL-11) production from the important model animals, mouse and macaque. The purified mouse and macaque rIL-11 interact with extracellular domain of human IL-11 receptor subunit α and activate STAT3 signaling in HEK293 cells co-expressing human IL-11 receptors with efficacies resembling those of human rIL-11. Hence, the evolutionary divergence does not impair IL-11 signaling. Furthermore, compared to human rIL-11 its macaque orthologue is 8-fold more effective STAT3 activator, which favors its use for treatment of thrombocytopenia as a potent substitute for human rIL-11. Compared to IL-6, IL-11 signaling exhibits lower species specificity, likely due to less conserved intrinsic disorder propensity within IL-6 orthologues. The developed express method for preparation of functionally active macaque/mouse rIL-11 samples is suited for exploration of the molecular mechanisms underlying IL-11 action and for development of the drug candidates for therapy of oncologic/hematologic/inflammatory diseases related to IL-11 signaling.
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. 212sok@gmail.com.
authors.forename: Andrei S
authors.initials: AS
authors.lastname: Sokolov
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. fenixfly@yandex.ru.
authors.forename: Alexei S
authors.initials: AS
authors.lastname: Kazakov
authors.affiliation: Biocad, Lubuchany 142380, Moscow Region, Russia. cvv@rambler.ru.
authors.forename: Valery V
authors.initials: VV
authors.lastname: Solovyev
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. ismailov_ramis@mail.ru.
authors.forename: Ramis G
authors.initials: RG
authors.lastname: Ismailov
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. vuversky@health.usf.edu.|Department of Molecular Medicine and USF Health Byrd Alzheimer's Research Institute, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA. vuversky@health.usf.edu.
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Uversky
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. lapteva.julia@gmail.com.
authors.forename: Yulia S
authors.initials: YS
authors.lastname: Lapteva
authors.affiliation: Antherix, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. romanuel@rambler.ru.
authors.forename: Roman V
authors.initials: RV
authors.lastname: Mikhailov
authors.affiliation: Antherix, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. catarios-ksu@mail.ru.
authors.forename: Ekaterina V
authors.initials: EV
authors.lastname: Pavlova
authors.affiliation: Antherix, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. ms.yterletskaya@mail.ru.
authors.forename: Iana O
authors.initials: IO
authors.lastname: Terletskaya
authors.affiliation: JSC R-Pharm, Berzarina str., 19/1, Moscow 123154, Russia. lv.ermolina@rpharm.ru.
authors.forename: Ludmila V
authors.initials: LV
authors.lastname: Ermolina
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. permyakov.s@gmail.com.
authors.forename: Sergei E
authors.initials: SE
authors.lastname: Permyakov
authors.affiliation: Institute for Biological Instrumentation of the Russian Academy of Sciences, Institutskaya str., 7, Pushchino 142290, Moscow Region, Russia. epermyak@yandex.ru.
authors.forename: Eugene A
authors.initials: EA
authors.lastname: Permyakov
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D017370:Interleukin-11 / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D008252:Macaca fascicularis; D051379:Mice; D053702:Receptors, Interleukin-11 / Q000378:metabolism*; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000235:genetics; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013045:Species Specificity
chemical_list: D017370:Interleukin-11; D015850:Interleukin-6; D053702:Receptors, Interleukin-11; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor
keywords: STAT3 signaling; bacterial expression; cloning; cytokines; interleukin-11; ligand-receptor interaction; protein-protein interaction; ubiquitin