The Disordered Structural Ensembles of Vasopressin and Oxytocin and Their Mutants

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1 The Disordered Structural Ensembles of Vasopressin and Oxytocin and Their Mutants Eugene Yedvabny, Paul S. Nerenberg 4, Clare So and Teresa Head-Gordon,,3* Department of Chemistry, Department of Bioengineering, 3 Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA USA 4 W.M. Keck Science Department, Claremont McKenna, Pitzer, and Scripps Colleges, Claremont, CA 7-576, USA.California SUPPLEMENTARY MATERIAL Table S-: Dominant hydrogen bonds for sub-ensembles of native oxytocin. Tyr -O Asn 5 -NH 73.% Tyr -O Cys 6 -NH 6.% Å Cys 6 -O Gly -NH 4.% Gln 4 -OE Gln 4 -NH 5.% Pro 7 -O C terminus-nh 4.3% Å Tyr -O Asn 5 -NH.6% Tyr -O Cys 6 -NH 7.6% Table S-: Dominant hydrogen bonds for sub-ensembles of Q4T mutant of oxytocin. Tyr -O Asn 5 -NH 5.% Tyr -O Cys 6 -NH 4.% Å Cys 6 -O Gly -NH 7.% Pro 7 -O C terminus-nh.3% Cys -O C terminus-nh.7% Å Tyr -O Asn 5 -NH 7.5% Tyr -O Cys 6 -NH 3.5% S

2 Table S-3: Dominant hydrogen bonds for sub-ensembles of Q4T,P7G mutant of oxytocin. Tyr -O Asn 5 -NH 7.6% Å Tyr -O Cys 6 -NH 7.7% Cys 6 -O Gly -NH 7.% Asn 5 -OD Tyr -NH 0.% Table S-4: Dominant hydrogen bonds for sub-ensembles of native vasopressin. Tyr -O Asn 5 -NH.% Tyr -O Cys 6 -NH 5.6% Å Pro 7 -O C terminus-nh 36.3% Cys 6 -O Gly -NH 3.% Cys -O C terminus-nh.% Pro 7 -O Gly -NH.0% Å Å Tyr -O Asn 5 -NH 0.7% Tyr -O Cys 6 -NH 63.5% Tyr -O Asn 5 -NH.7% Tyr -O Cys 6 -NH 4.5% Phe 3 -O Cys 6 -NH 7.4% Table S-5: Dominant hydrogen bonds for sub-ensembles of YH mutant of vasopressin. His -O Asn 5 -NH 7.5% His -O Cys 6 -NH 76.7% Å His -ND Gln 4 -NH 7.6% Pro 7 -O C terminus-nh 33.4% Cys 6 -O Gly -NH 30.7% Cys -O C terminus-nh.% Å His -O Asn 5 -NH 0.6% His -O Cys 6 -NH 7.7% His -ND Gln 4 -NH 0.% S

3 Table S-6: Dominant hydrogen bonds for sub-ensembles of YH+ mutant of vasopressin. His -O Asn 5 -NH 74.6% His -O Cys 6 -NH 5.6% Å His -O C terminus-nh 6.6% Cys 6 -O Gly -NH 5.3% Gly -O Cys 6 -NH 54.% Asn 5 -OD His -HD.% Å His -O Asn 5 -NH 6.4% His -O Cys 6 -NH.% Asn 5 -OD His -HD 35.7% Table S-7: Dominant hydrogen bonds for sub-ensembles of P7L mutant of vasopressin. Tyr -O Asn 5 -NH 3.% Tyr -O Cys 6 -NH 6.0% Asn 5 -OD Leu 7 -NH.% Gln 4 -O Arg -NH.% Å Phe 3 -O Cys 6 -NH.% Asn 5 -O Arg -NH 7.5% Phe 3 -O Leu 7 -NH 7.0% Tyr -O Cys 6 -NH 6.0% Phe 3 -O Asn 5 -NH 3.44% Gln 4 -O Leu 7 -NH.4% Å Å Tyr -O Asn 5 -NH 4.4% Gln 4 -O Leu 7 -NH 6.3% Phe 3 -O Cys 6 -NH.% Asn 5 -O Arg -NH.0% Tyr -O Asn 5 -NH 44.% S3

4 Table S-: Percentages of sub-ensembles of native oxytocin with specific DSSP secondary structure. Blank entries imply 0.0%. R g bounds Residue β-bridge 3 0 helix Turn 3.6% 3.6% 4.6% 5.% Å 5.6% 4.% 6.6% % 67.5%.6% Å 3 3.6%.0% 4 3.6%.% 5 3.6%.% 6 7.0% Table S-: Percentages of sub-ensembles of Q4T mutant of oxytocin with specific DSSP R g bounds Residue 3 0 helix Turn 3.%.3% 4.%.4% Å 5.%.4% 6.% 7 0.4% 0.5% Å 3.6% 5.4% 4.6% 5.% 5.6% 5.% 6 7 S4

5 Table S-3: Percentages of sub-ensembles of Q4T,P7G mutant of oxytocin with specific DSSP R g bounds Residue 3 0 helix α-helix Turn 3 4.3%.% 7.7% 4 4.3%.% 0.0% Å 5 4.3%.% 7.3% 6.% 3.0% 7.%.0% Table S-4: Percentages of sub-ensembles of native vasopressin with specific DSSP secondary structure. Blank entries imply 0.0%. R g bounds Residue 3 0 helix Turn 3.7%.0% 4.7%.0% Å 5.7% 7.0% 6.% % 6.% Å Å 3.3%.6% 4.3%.7% 5.3%.% 6.0% 7.0% 3 3.% 76.% 4 3.% 76.% 5 3.% 7.5% 6 7 S5

6 Table S-5: Percentages of sub-ensembles of YH mutant of vasopressin with specific DSSP R g bounds Residue 3 0 helix Turn 3.0%.% 4.0%.% Å 5.0%.0% 6 3.0% 7 6.4% 6.5% Å 3.6% 7.7% 4.6% 7.6% 5.6% 6.4% 6.6% 7.6% Table S-6: Percentages of sub-ensembles of YH+ mutant of vasopressin with specific DSSP Parallel Anti-parallel R g bounds Residue 3 β-bridge β-bridge 0 helix Turn Å Å 3 7.%.% 6.7% 4.% 7.4% 5.% 55.0% 6 7.% 5.0% % 77.3% 5.0%.%.6% 3.0% 3.4% 3.% 4 3.4% 0.% 5.%.0% 5.5% 6.0% 7 S6

7 Table S-7: Percentages of sub-ensembles of P7L mutant of vasopressin with specific DSSP R g bounds Å Å Å Residue Parallel β-bridge Anti-parallel β-bridge 3 0 helix α-helix Turn 7.% 3.5% 3.% 3.5%.0% 47.% 4 5.3% 4.% 60.3% 5 7.6% 4.0%.4% 4.5% 6.% 4.3%.4% 34.6% 7 3.%.7% 34.7%.% 5.7%.0%.0% 6.0% 3.0% 6.% 47.3% 4.%.6% 76.7% 5.0% 6.%.% 66.7% 6.0% 7.3%.% 4.3% 7.0%.7%.0%.6% 0.4% 3.%.0% 3.3% 57.4% 4.6%.5% 5 3.%.% 6.3% 6.%.% 7.0% 7.7% S7

8 Table S3-: The experimental and simulated chemicals shifts for native Oxytocin. Experimental values taken from Ohno et al. C α C β H α H N Residue Exp Sim Exp Sim Exp Sim Exp Sim Table S3-: The experimental and simulated chemicals shifts for native Oxytocin. Experimental values taken from Sikorska et al. 3 C α C β H α H N Residue Exp Sim Exp Sim Exp Sim Exp Sim S

9 Figure S. Difference between the simulated R g distributions of native oxytocin for two independent 45 ns production runs, showing the convergence of our simulations. Oxytocin R g Convergence Profile χ Simulation Time (ns) REFERENCES. Kabsch, W.; Sander, C., Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features. Biopolymers 3,, Ohno, A.; Kawasaki, N.; Fukuhara, K.; Okuda, H.; Yamaguchi, T., Complete NMR analysis of oxytocin in phosphate buffer. Magnetic Resonance in Chemistry 00, 4, Sikorska, E.; Rodziewicz-Motowidło, S., Conformational studies of vasopressin and mesotocin using NMR spectroscopy and molecular modelling methods. Part I: Studies in water. Journal of peptide science : an official publication of the European Peptide Society 00, 4, S