Decoupling of size and shape fluctuations in heteropolymeric sequences reconciles discrepancies in SAXS vs. FRET measurements.

Fuertes G, Banterle N, Ruff KM, Chowdhury A, Mercadante D, Koehler C, Kachala M, Estrada Girona G, Milles S, Mishra A, Onck PR, Gräter F, Esteban-Martín S, Pappu RV, Svergun DI, Lemke EA
Proc Natl Acad Sci U S A 114(31):E6342-E6351 (2017 Aug 1)

PMID: 28716919
doi: 10.1073/pnas.1704692114

Submitted to SASBDB: 2018 Aug 16
Published in SASBDB: 2018 Aug 18

SASDEH2 – Unlabeled nucleoporin NUP49/NSP49 (N49) without denaturant

Nucleoporin NUP49/NSP49 experimental SAS data

Unlabeled nucleoporin NUP49/NSP49 (N49) without denaturant Rg histogram

Sample:	Nucleoporin NUP49/NSP49 monomer, 4 kDa Saccharomyces cerevisiae protein
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Dec 8

R_g^Guinier	1.6	nm
D_max	6.0	nm
Volume^Porod	4	nm³

SASDEJ2 – Labeled nucleoporin NUP49/NSP49 (N49-Alexa488/Alexa594) without denaturant

Nucleoporin NUP49/NSP49Alexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Nucleoporin NUP49/NSP49 Alexa Fluor™ 594 C5 Maleimide Alexa Fluor™ 488 C5 Hydroxylamine Kratky plot

Sample:	Nucleoporin NUP49/NSP49 monomer, 4 kDa Saccharomyces cerevisiae protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Dec 8

R_g^Guinier	1.9	nm
D_max	6.5	nm
Volume^Porod	12	nm³

SASDEK2 – Unlabeled nucleoporin NUP49/NSP49 (N49) with denaturant

Unlabeled nucleoporin NUP49/NSP49 (N49) with denaturant Rg histogram

Sample:	Nucleoporin NUP49/NSP49 monomer, 4 kDa Saccharomyces cerevisiae protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Dec 8

R_g^Guinier	1.7	nm
D_max	6.5	nm
Volume^Porod	4	nm³

SASDEL2 – Labeled nucleoporin NUP49/NSP49 (N49-Alexa488/Alexa594) with denaturant

Sample:	Nucleoporin NUP49/NSP49 monomer, 4 kDa Saccharomyces cerevisiae protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	2.1	nm
D_max	7.7	nm
Volume^Porod	7	nm³

SASDEM2 – Unlabeled Nuclear Localization Signal (NLS) from the inner nuclear membrane protein HEH2 without denaturant

Inner nuclear membrane protein HEH2 experimental SAS data

Unlabeled Nuclear Localization Signal (NLS) from the inner nuclear membrane protein HEH2 without denaturant Rg histogram

Sample:	Inner nuclear membrane protein HEH2 monomer, 5 kDa Saccharomyces cerevisiae protein
Buffer:	25 mM HEPES, 150 mM NaCl, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2014 Jan 25

R_g^Guinier	2.4	nm
D_max	11.6	nm
Volume^Porod	16	nm³

SASDEN2 – Labeled Nuclear Localization Signal from the inner nuclear membrane protein HEH2 (NLS-Alexa488/Alexa594) without denaturant

Inner nuclear membrane protein HEH2Alexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Inner nuclear membrane protein HEH2 Alexa Fluor™ 594 C5 Maleimide Alexa Fluor™ 488 C5 Hydroxylamine Kratky plot

Sample:	Inner nuclear membrane protein HEH2 monomer, 5 kDa Saccharomyces cerevisiae protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	25 mM HEPES, 150 mM NaCl, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2014 Jan 25

R_g^Guinier	2.0	nm
D_max	7.3	nm
Volume^Porod	10	nm³

SASDEP2 – Unlabeled Nuclear Localization Signal (NLS) from the inner nuclear membrane protein HEH2 with denaturant

Unlabeled Nuclear Localization Signal (NLS) from the inner nuclear membrane protein HEH2 with denaturant Rg histogram

Sample:	Inner nuclear membrane protein HEH2 monomer, 5 kDa Saccharomyces cerevisiae protein
Buffer:	25 mM HEPES, 150 mM NaCl, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2014 Jan 25

R_g^Guinier	2.3	nm
D_max	7.1	nm
Volume^Porod	10	nm³

SASDEQ2 – Labeled Nuclear Localization Signal from the inner nuclear membrane protein HEH2 (NLS-Alexa488/Alexa594) with denaturant

Sample:	Inner nuclear membrane protein HEH2 monomer, 5 kDa Saccharomyces cerevisiae protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	25 mM HEPES, 150 mM NaCl, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2014 Jan 25

R_g^Guinier	2.4	nm
D_max	8.1	nm
Volume^Porod	12	nm³

SASDER2 – Unlabeled Importin Beta Binding domain (IBB) from importin subunit alpha-1 without denaturant

Importin subunit alpha-1 experimental SAS data

Unlabeled Importin Beta Binding domain (IBB) from importin subunit alpha-1 without denaturant Rg histogram

Sample:	Importin subunit alpha-1 monomer, 11 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Dec 8

R_g^Guinier	3.2	nm
D_max	11.5	nm
Volume^Porod	30	nm³

SASDES2 – Labeled Importin Beta Binding Domain (IBB-Alexa488/Alexa594) from importin subunit alpha-1 without denaturant

Importin subunit alpha-1Alexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Importin subunit alpha-1 Alexa Fluor™ 594 C5 Maleimide Alexa Fluor™ 488 C5 Hydroxylamine Kratky plot

Sample:	Importin subunit alpha-1 monomer, 11 kDa Homo sapiens protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Dec 9

R_g^Guinier	2.9	nm
D_max	13.7	nm
Volume^Porod	29	nm³

SASDET2 – Unlabeled Importin Beta Binding Domain (IBB) from importin subunit alpha-1 with denaturant

Unlabeled Importin Beta Binding Domain (IBB) from importin subunit alpha-1 with denaturant Rg histogram

Sample:	Importin subunit alpha-1 monomer, 11 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	3.1	nm
D_max	15.3	nm
Volume^Porod	34	nm³

SASDEU2 – Labeled Importin Beta Binding Domain from importin subunit alpha-1 (IBB-Alexa488/Alexa594) with denaturant

Sample:	Importin subunit alpha-1 monomer, 11 kDa Homo sapiens protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	3.2	nm
D_max	13.3	nm
Volume^Porod	29	nm³

SASDEV2 – Unlabeled nuclear pore complex protein Nup153 (NUS) without denaturant

Nuclear pore complex protein Nup153 experimental SAS data

Unlabeled nuclear pore complex protein Nup153 (NUS) without denaturant Rg histogram

Sample:	Nuclear pore complex protein Nup153 monomer, 8 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	2.5	nm
D_max	9.2	nm
Volume^Porod	15	nm³

SASDEW2 – Labeled nuclear pore complex protein Nup153 (NUS-Alexa488/Alexa594) without denaturant

Nuclear pore complex protein Nup153Alexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Nuclear pore complex protein Nup153 Alexa Fluor™ 594 C5 Maleimide Alexa Fluor™ 488 C5 Hydroxylamine Kratky plot

Sample:	Nuclear pore complex protein Nup153 monomer, 8 kDa Homo sapiens protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	2.5	nm
D_max	9.7	nm
Volume^Porod	17	nm³

SASDEX2 – Unlabeled nuclear pore complex protein Nup153 (NUS) with denaturant

Unlabeled nuclear pore complex protein Nup153 (NUS) with denaturant Rg histogram

Sample:	Nuclear pore complex protein Nup153 monomer, 8 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	3.1	nm
D_max	11.0	nm
Volume^Porod	26	nm³

SASDEY2 – Labeled nuclear pore complex protein Nup153 (NUS-Alexa488/Alexa594) with denaturant

Sample:	Nuclear pore complex protein Nup153 monomer, 8 kDa Homo sapiens protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	2.9	nm
D_max	10.8	nm
Volume^Porod	19	nm³

SASDEZ2 – Unlabeled nuclear pore complex protein Nup153 (NUL) without denaturant

Unlabeled nuclear pore complex protein Nup153 (NUL) without denaturant Rg histogram

Sample:	Nuclear pore complex protein Nup153 monomer, 12 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	3.0	nm
D_max	10.9	nm
Volume^Porod	32	nm³

SASDE23 – Labeled nuclear pore complex protein Nup153 (NUL-Alexa488/Alexa594) without denaturant

Sample:	Nuclear pore complex protein Nup153 monomer, 12 kDa Homo sapiens protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	3.1	nm
D_max	12.0	nm
Volume^Porod	24	nm³

SASDE33 – Unlabeled nuclear pore complex protein Nup153 (NUL) with denaturant

Unlabeled nuclear pore complex protein Nup153 (NUL) with denaturant Rg histogram

Sample:	Nuclear pore complex protein Nup153 monomer, 12 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	3.5	nm
D_max	16.0	nm
Volume^Porod	53	nm³

SASDE43 – Labeled nuclear pore complex protein Nup153 (NUL-Alexa488/Alexa594) with denaturant

Sample:	Nuclear pore complex protein Nup153 monomer, 12 kDa Homo sapiens protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	3.2	nm
D_max	12.0	nm
Volume^Porod	27	nm³

SASDE53 – Unlabeled dihydrolipoyllysine-residue succinyltransferase component (BBL) with denaturant

Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex experimental SAS data

Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex Kratky plot

Sample:	Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex monomer, 4 kDa Escherichia coli protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	2.1	nm
D_max	8.3	nm
Volume^Porod	41	nm³

SASDE63 – Labeled dihydrolipoyllysine-residue succinyltransferase component (BBL-Alexa488/Alexa594) with denaturant

Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complexAlexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Sample:	Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex monomer, 4 kDa Escherichia coli protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Nov 8

R_g^Guinier	2.3	nm
D_max	10.7	nm
Volume^Porod	13	nm³

SASDE73 – Unlabeled cold shock protein (CSP) with denaturant

Cold shock-like protein experimental SAS data

Sample:	Cold shock-like protein monomer, 7 kDa Thermotoga maritima protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Mar 16

R_g^Guinier	2.5	nm
D_max	11.5	nm
Volume^Porod	17	nm³

SASDE83 – Labeled cold shock protein (CSP-Alexa488/Alexa594) with denaturant

Cold shock-like proteinAlexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Cold shock-like protein Alexa Fluor™ 594 C5 Maleimide Alexa Fluor™ 488 C5 Hydroxylamine Kratky plot

Sample:	Cold shock-like protein monomer, 7 kDa Thermotoga maritima protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	2.2	nm
D_max	8.3	nm
Volume^Porod	9	nm³

SASDE93 – Unlabeled thioredoxin (TRX) with denaturant

Sample:	Thioredoxin 1 monomer, 12 kDa Escherichia coli protein
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	3.6	nm
D_max	13.0	nm
Volume^Porod	34	nm³

SASDEA3 – Labeled thioredoxin (TRX-Alexa488/Alexa594) with denaturant

Thioredoxin 1Alexa Fluor™ 594 C5 MaleimideAlexa Fluor™ 488 C5 Hydroxylamine experimental SAS data

Thioredoxin 1 Alexa Fluor™ 594 C5 Maleimide Alexa Fluor™ 488 C5 Hydroxylamine Kratky plot

Sample:	Thioredoxin 1 monomer, 12 kDa Escherichia coli protein Alexa Fluor™ 594 C5 Maleimide monomer, 1 kDa Alexa Fluor™ 488 C5 Hydroxylamine monomer, 1 kDa
Buffer:	PBS, 10 mM DTT, 6 M urea, 0.3 M KCl, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2013 Jun 15

R_g^Guinier	3.2	nm
D_max	13.9	nm
Volume^Porod	31	nm³

SASDEB3 – Unlabeled nuclear pore complex protein Nup98-Nup96 (N98) without denaturant

Nuclear pore complex protein Nup98-Nup96 experimental SAS data

Nuclear pore complex protein Nup98-Nup96 Kratky plot

Sample:	Nuclear pore complex protein Nup98-Nup96 monomer, 15 kDa Homo sapiens protein
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2015 Jun 24

R_g^Guinier	2.9	nm
D_max	10.4	nm
Volume^Porod	27	nm³

SASDEC3 – Unlabeled nucleoporin NSP1 (NSP) without denaturant

Sample:	Nucleoporin NSP1 monomer, 18 kDa Saccharomyces cerevisiae protein
Buffer:	PBS, 10 mM DTT, pH: 7.4
Experiment:	SAXS data collected at EMBL P12, PETRA III on 2015 Jun 24

R_g^Guinier	4.1	nm
D_max	15.0	nm
Volume^Porod	45	nm³