Surface electrostatics dictate RNA-binding protein CAPRIN1 condensate concentration and hydrodynamic properties

Abstract

Biomolecular condensates concentrate proteins, nucleic acids, and small molecules and play an essential role in many biological processes. Their formation is tuned by a balance between energetically favorable and unfavorable contacts, with charge–charge interactions playing a central role in some systems. The positively charged intrinsically disordered carboxy-terminal region of the RNA-binding protein CAPRIN1 is one such example, phase separating upon addition of negatively charged ATP or high concentrations of sodium chloride (NaCl). Using solution NMR spectroscopy, we measured residue-specific near-surface electrostatic potentials (ϕENS) of CAPRIN1 along its NaCl-induced phase separation trajectory to compare with those obtained using ATP. In both cases, electrostatic shielding decreases ϕENS values, yet surface potentials of CAPRIN1 in the two condensates can be different, depending on the amount of NaCl or ATP added. Our results establish that even small differences in ϕENS can significantly affect the level of protein enrichment and the mechanical properties of the condensed phase, leading, potentially, to the regulation of biological processes. Biomolecular condensates concentrate proteins, nucleic acids, and small molecules and play an essential role in many biological processes. Their formation is tuned by a balance between energetically favorable and unfavorable contacts, with charge–charge interactions playing a central role in some systems. The positively charged intrinsically disordered carboxy-terminal region of the RNA-binding protein CAPRIN1 is one such example, phase separating upon addition of negatively charged ATP or high concentrations of sodium chloride (NaCl). Using solution NMR spectroscopy, we measured residue-specific near-surface electrostatic potentials (ϕENS) of CAPRIN1 along its NaCl-induced phase separation trajectory to compare with those obtained using ATP. In both cases, electrostatic shielding decreases ϕENS values, yet surface potentials of CAPRIN1 in the two condensates can be different, depending on the amount of NaCl or ATP added. Our results establish that even small differences in ϕENS can significantly affect the level of protein enrichment and the mechanical properties of the condensed phase, leading, potentially, to the regulation of biological processes. The phase separation of biomolecules controls many aspects of cellular function (1Hyman A.A. Weber C.A. Jülicher F. Liquid-liquid phase separation in biology.Annu. Rev. Cell Dev. Biol. 2014; 30: 39-58Crossref PubMed Scopus (1664) Google Scholar, 2Banani S.F. Lee H.O. Hyman A.A. Rosen M.K. Biomolecular condensates: organizers of cellular biochemistry.Nat. Rev. Mol. Cell Biol. 2017; 18: 285-298Crossref PubMed Scopus (2593) Google Scholar), and understanding the atomic details of the driving forces underlying this process has, therefore, been the subject of considerable efforts. The interactions that regulate the formation and disassembly of biological condensates have been extensively characterized via biochemical and computational studies (2Banani S.F. Lee H.O. Hyman A.A. Rosen M.K. Biomolecular condensates: organizers of cellular biochemistry.Nat. Rev. Mol. Cell Biol. 2017; 18: 285-298Crossref PubMed Scopus (2593) Google Scholar, 3Wang J. Choi J.-M. Holehouse A.S. Lee H.O. Zhang X. Jahnel M. et al.A molecular grammar governing the driving forces for phase separation of prion-like RNA binding proteins.Cell. 2018; 174: 688-699.e16Abstract Full Text Full Text PDF PubMed Scopus (893) Google Scholar, 4Martin E.W. Holehouse A.S. Peran I. Farag M. Incicco J.J. Bremer A. et al.Valence and patterning of aromatic residues determine the phase behavior of prion-like domains.Science. 2020; 367: 694-699Crossref PubMed Scopus (380) Google Scholar); however, site-specific experimental information is lacking. Among the various interactions, the role of electrostatics is of particular interest as phase-separating proteins can be enriched in charged residues (5Chong P.A. Vernon R.M. Forman-Kay J.D. RGG/RG motif regions in RNA binding and phase separation.J. Mol. Biol. 2018; 430: 4650-4665Crossref PubMed Scopus (201) Google Scholar, 6Das R.K. Pappu R.V. Conformations of intrinsically disordered proteins are influenced by linear sequence distributions of oppositely charged residues.Proc. Natl. Acad. Sci. U. S. A. 2013; 110: 13392-13397Crossref PubMed Scopus (557) Google Scholar), and altering protein surface electrostatics by interactions with charged molecules such as ATP or RNA (7Patel A. Malinovska L. Saha S. Wang J. Alberti S. Krishnan Y. et al.ATP as a biological hydrotrope.Science. 2017; 356: 753-756Crossref PubMed Scopus (508) Google Scholar, 8Maharana S. Wang J. Papadopoulos D.K. Richter D. Pozniakovsky A. 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Hein M.Y. et al.A liquid-to-solid phase transition of the ALS protein FUS accelerated by disease mutation.Cell. 2015; 162: 1066-1077Abstract Full Text Full Text PDF PubMed Scopus (1575) Google Scholar, 12Monahan Z. Ryan V.H. Janke A.M. Burke K.A. Rhoads S.N. Zerze G.H. et al.Phosphorylation of the FUS low-complexity domain disrupts phase separation, aggregation, and toxicity.EMBO J. 2017; 36: 2951-2967Crossref PubMed Scopus (415) Google Scholar) can dramatically affect a protein’s phase-separation propensity both in vitro and in cells. One prominent example where electrostatic interactions are critical to phase separation is provided by the 709 residue protein CAPRIN1, a molecule found in membraneless organelles, such as stress granules, P bodies, and mRNA transport granules that play important roles in regulating RNA processing (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar, 14Jiang Y. X. Wang M. X. et of in by J. 2013; Full Text Full Text PDF PubMed Scopus Google have that the low-complexity region of CAPRIN1 to as phase in and its small NMR studies of the and interactions that its phase separation B. Vernon R.M. Kay L.E. Forman-Kay J.D. phase separation of and CAPRIN1 regulation of and PubMed Scopus Google Scholar, L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google Scholar, Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; CAPRIN1 two the and with a region enriched in aromatic residues CAPRIN1 a of and a of phase separation upon of unfavorable electrostatic interactions between positively charged CAPRIN1 in vitro this can be the addition of charged molecules such as ATP or RNA or by the addition of high concentrations of sodium chloride for a CAPRIN1 B. Vernon R.M. Kay L.E. Forman-Kay J.D. phase separation of and CAPRIN1 regulation of and PubMed Scopus Google Scholar, L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google Scholar, Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; In to electrostatics regulate a particular biological a of the surface electrostatic potentials of the molecular is is by the using of the proteins of interest B. A. electrostatics in and PubMed Scopus Google Scholar, F. A. The for a for Mol. PubMed Scopus Google the of such a computational is with for intrinsically disordered proteins such as CAPRIN1 that in solution NMR an for the experimental of per-residue near-surface electrostatic potentials (ϕENS) in such Y. A. of for 2020; PubMed Scopus Google Scholar, B. J. of near-surface electrostatic potentials by Natl. Acad. Sci. U. S. A. 2021; Scopus Google Scholar, B. J. electrostatics NMR for B. 2022; PubMed Scopus Google The are on to of NMR with of ϕENS a of have the to ϕENS of CAPRIN1 along its phase separation trajectory as concentrations of ATP are to phase separation (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google In a in CAPRIN1 phase ϕENS are and to with the and regions of the addition of small of ATP ϕENS to the binding of ATP to the and regions that to of as a in the phase separation process (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google In the condensed phase, that ATP to the condensed to high concentrations of protein and is with ATP molecules with one CAPRIN1 concentrations the binding of ATP to the of CAPRIN1 to small potentials in regions and is by of condensates and into a between in electrostatic potential and interactions, the region of CAPRIN1, (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google we have that the addition of NaCl phase separation of CAPRIN1, a for a CAPRIN1 L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google Scholar) with ATP Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; is to that the addition of unfavorable electrostatic interactions between CAPRIN1 in a to ATP. we the ϕENS of CAPRIN1 in the of NaCl that of ATP the of of the two with CAPRIN1 are example, ATP can with the regions and with the process Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; Scholar, Y. The role of ATP in RNA protein in 2022; PubMed Scopus (8) Google Scholar) interactions are by between on E.W. Holehouse A.S. Peran I. Farag M. Incicco J.J. Bremer A. et al.Valence and patterning of aromatic residues determine the phase behavior of prion-like domains.Science. 2020; 367: 694-699Crossref PubMed Scopus (380) Google Scholar, Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; Scholar) that be by the binding of ATP. high concentrations of NaCl are to favorable interactions G. S. E. et liquid phase of proteins is by and 2021; PubMed Scopus Google the of for CAPRIN1 phase separation with NaCl and the of the interactions that are be with ATP. this is the is that the properties of the we have CAPRIN1 ϕENS as a function of NaCl and characterized the interactions and of CAPRIN1 in the condensed differences found between the electrostatic potentials of and in to differences in In to the of NaCl for phase separation of a CAPRIN1 we as a function of The in that formation that NaCl concentrations in of are with results that high concentrations of are to phase separation L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google measured per-residue ϕENS NaCl and where CAPRIN1 is in the phase to establish ϕENS the phase separation the by et B. J. of near-surface electrostatic potentials by Natl. Acad. Sci. U. S. A. 2021; Scopus Google Scholar), and as in (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar), of measured using the charged and ϕENS the B. J. of near-surface electrostatic potentials by Natl. Acad. Sci. U. S. A. 2021; Scopus Google where is is the where and and are the measured and is the of the of an on we NMR of CAPRIN1 in the of NaCl and in the of The small and in the and with region that the is a of in surface electrostatics by of per-residue ϕENS measured a of NaCl and the NaCl ϕENS the of by the B. J. of the of the electrostatic potential of proteins in and B. 2022; PubMed Scopus Google where CAPRIN1 to phase ϕENS are small on that is for CAPRIN1 phase separation, as with ATP (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google in even concentrations of to of CAPRIN1 molecules in the a of CAPRIN1, in NMR studies of phase phase separation NaCl concentrations to a in the protein that is in NMR as the condensed phase that to the of the NMR protein phase ϕENS high therefore, a of CAPRIN1, where the residues are with to significantly the phase separation propensity the of the as L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google In the of ϕENS of CAPRIN1 and CAPRIN1 in phase are compare and in the of the to on is that as the CAPRIN1 surface the electrostatic interactions between are we on to NMR molecules and the on in molecules using a of and by mutations and with the of the via a and are in the region the and the region of CAPRIN1, both of are in CAPRIN1 that are important in the (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar, Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; differences in measured for the and CAPRIN1, and CAPRIN1 by to the the and measured in the or the of In the of the electrostatic between CAPRIN1 that In significantly measured with interactions, that the on a and the on an in the of high The the two are with two in the of the with results studies on measured in the condensed Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; results as (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar), that the aromatic regions are for that to the of the and that the interactions are regions of that the are to those measured in the of ATP (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar), that regions with and the of the between and results that the for formation are between and phase is important to that the ϕENS on The small amount of that is NaCl decreases the of the the electrostatic potential measured to the of phase separation, is the potential condensed phase In to residue-specific ϕENS of CAPRIN1 molecules in the condensed phase, we have a a NaCl of for the of and interactions in the condensed phase Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; that for the high concentrations of protein to is in to CAPRIN1 concentrations where is liquid in the NMR a phase on in and a phase on the or with a (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar, Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; Scholar), is to that the condensed phase is by the region of the in the NMR with the phase the an G. by of and an to NMR of biological in Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar) of CAPRIN1 in the condensed phase using a protein in an in the of and ϕENS such with and a or The of ϕENS obtained with those measured using the various concentrations of NaCl or as as the condensed phase (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar) The ϕENS the condensed phase significantly those measured in the in the of the electrostatic shielding by the in the however, the ϕENS between and that CAPRIN1 is positively charged in the condensed per-residue electrostatic potentials the is in to the condensed phase where CAPRIN1 negatively charged to with the of the condensed phase upon addition of ATP (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google in the in the surface electrostatic potentials between the and condensed to in the of CAPRIN1 and in the properties of the this we NMR to the of CAPRIN1 in the and condensed The of CAPRIN1 and in the and condensed to that are in the ATP a of to properties in separating the potential of properties D. Z. S. Y. et of an NMR to the of a with PubMed Scopus Google the function where the and the of the of on the D. Z. S. Y. et of an NMR to the of a with PubMed Scopus Google Scholar, Zhang A. Kay L.E. function using PubMed Scopus Google The are in the condensed phase, that the of of CAPRIN1 is significantly to the in the of between the two condensed phase have that the site-specific in the two regions results that are for the of the (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar, Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; the between that a of residues is in the formation of CAPRIN1 in both condensed phase is of interest to that the is significantly the between and condensed that molecular is that the concentrations of protein in condensed are the the transition a to a solution to the where protein molecules for CAPRIN1 molecules with as significantly of the protein or are is the of the CAPRIN1 properties in the two condensed The of CAPRIN1 in the condensed phase by addition of ATP a of is (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar), that measured in the condensed phase The of CAPRIN1 in the droplets to the of protein and that the amount of CAPRIN1 in the condensed phase in to the of electrostatic between protein molecules as by ϕENS In this the potential in the the of CAPRIN1 molecules to condensed where CAPRIN1 is and interactions, this we measured the CAPRIN1 in a with a of NaCl where ϕENS are to be to as measured for the in the the of CAPRIN1 significantly to to the measured in the is in with phase that we have in the condensed phase CAPRIN1 significantly with the of Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; that the CAPRIN1 electrostatic potential the protein the properties of the and that the of of ϕENS on the of of the with ATP chloride of its the results of this CAPRIN1 in a and surface electrostatic potential the molecule with the and regions of the protein charge–charge addition of of the surface potential of CAPRIN1 to interactions to phase The per-residue ϕENS in the condensed phase using NaCl establish that CAPRIN1 is in to droplets with where of CAPRIN1 is (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Our results in the and the in the condensed phase that the interactions for the are between the two condensed with the of residues in the formation of differences in protein concentrations in the and ATP condensates differences in of CAPRIN1 molecules in affect the properties of the Our that even small in electrostatic potentials of a can to in the properties of the condensed phase is CAPRIN1 with RNA and binding such as protein that can be and its can be via post-translational modifications B. Vernon R.M. Kay L.E. Forman-Kay J.D. phase separation of and CAPRIN1 regulation of and PubMed Scopus Google to surface electrostatics play an important role in critical concentrations of both and the as as the properties of molecules in the condensed phase, as we have for example, of The low-complexity region of CAPRIN1, residues to 709 and its where the in the sequence with and as (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar, B. Vernon R.M. Kay L.E. Forman-Kay J.D. phase separation of and CAPRIN1 regulation of and PubMed Scopus Google Scholar, L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google and can the of CAPRIN1 and of in the NMR L.E. Forman-Kay J.D. Kay L.E. NMR for studies of and condensed protein to the phase-separating protein 2020; PubMed Scopus Google we and mutations and the in the in this to as CAPRIN1 in and mutations and mutations and by using a have that the of the and mutations or to or molecules the phase separation of CAPRIN1 (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google NMR of CAPRIN1, and with or to The of CAPRIN1 to phase separation, of electrostatic potentials high as (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar, B. Vernon R.M. Kay L.E. Forman-Kay J.D. phase separation of and CAPRIN1 regulation of and PubMed Scopus Google separation by to CAPRIN1 protein with NaCl in and to the CAPRIN1 solution and to phase The and and a to concentrations of and to the condensed phase to the concentrations of as and in (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google the phase, to the condensed to a NMR on CAPRIN1 as (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google CAPRIN1 by with the by the solution using a with of in sodium and The by with of by using a The of the by CAPRIN1 with of The by an using a to the modifications using CAPRIN1 to a protein of with concentrations of NaCl to into a and using a the NMR on a or using a with and The NMR on a with a liquid and details are as (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google electrostatic potentials obtained with a of and using as B. J. of near-surface electrostatic potentials by Natl. Acad. Sci. U. S. A. 2021; Scopus Google Scholar), where or for the or In the residues with of of in using the of et B. J. of near-surface electrostatic potentials by Natl. Acad. Sci. U. S. A. 2021; Scopus Google have a for concentrations of in the of the in (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google differences in concentrations of for by of using where and are measured and and are the or are to the of the measured in the condensed phase is the of as a function of the of obtained into the that of or in be by one a (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google In this the of between and in the condensed phase by by The significantly the for the condensed phase in (13Toyama Y. Rangadurai A.K. Forman-Kay J.D. Kay L.E. Mapping the per-residue surface electrostatic potential of CAPRIN1 along its phase-separation trajectory.Proc. Natl. Acad. Sci. U. S. A. 2022; 119e2210492119Crossref Scopus (8) Google Scholar), of the between the the favorable of into the condensed The of CAPRIN1 in the condensed phase measured using the in the by et K.A. S. et of molecular an using and NMR Mol. Biol. PubMed Scopus Google Scholar) with a to a to aromatic ATP that with the of of with of or and for with to obtained for both that is an in NMR The The function where is the and is the of the in by for a of and and with in the of and on and along with and The as D. Z. S. Y. et of an NMR to the of a with PubMed Scopus Google Scholar), a between and of and an with to the of the to measured as J. A. of in proteins by PubMed Scopus Google Scholar) using to by in the and of J. A. of in proteins by PubMed Scopus Google Scholar, F. A. D. the of PubMed Scopus Google on G. by of and an to NMR of biological in Natl. Acad. Sci. U. S. A. PubMed Scopus Google that to to in the condensed phase and in the condensed Z. Y. Kay L.E. of in proteins by NMR PubMed Scopus Google the phase of CAPRIN1 by to CAPRIN1 with of a solution of to concentrations of or in concentrations in both and condensed obtained by the and of the condensed and into a and in of and as Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; The condensed phase solution using a Lee Y. Forman-Kay J.D. et for phase separation by NMR studies of a CAPRIN1 condensed Natl. Acad. Sci. U. S. A. 2021; that the of the critical for the of the condensed of CAPRIN1 in the condensed and are and for the with NaCl and and for the with and obtained by on the condensed phase and for and The the are the upon of CAPRIN1 have been in the The that have of interest with the of this for for L. E. Y. A. and L. E. Y. A. J. D. and L. E. Y. A. and L. E. Y. A. and L. E. Y. T. and L. E. Y. A. J. D. and L. E. Y. T. is a for the of an and a the of A. is to the for by L. E. J. D. L. E. J. D. and of J. D. J. D. is by the The is the of the and the of the of

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