European Biophysics Journal

Electronic Supplementary Material

Folding of an All-Helical Greek-Key Protein Monitored by Quenched-Flow Hydrogen-

Deuterium Exchange and NMR Spectroscopy

Lesley H. Greene1,* · Hai Li1 · Junyan Zhong2 · Guoxia Zhao1 · Khym Wilson1

1Department of Chemistry and Biochemistry, 4541 Hampton Boulevard, Old Dominion

University, Norfolk, VA 23529, USA; 2College of Sciences Major Instrumentation Cluster, 4402

Elkhorn Avenue, Old Dominion University, Norfolk, VA 23529, USA

*To whom correspondence should be addressed: L.H. Greene, Department of Chemistry and

Biochemistry, 4541 Hampton Boulevard, Old Dominion University, Norfolk, VA 23529, USA.

E-mail: [email protected]

Figure S1. The HSQC spectrum of Fadd-DD in 100 mM K2HPO4/50 mM citric acid, 5 mM DTT

(pH 4.8) and 10% D2O at 30°C. The twenty-four stable amide protons in our kinetic study are indicated in bold and underlined.

2 Figure S2. Simple schematic model of protein folding for Fadd-DD based on experimental data.

The helices are denoted with numbers 1-6. The hydrophobic core is depicted by the dotted circle. Here all six helices are formed concomitant with the establishment of the hydrophobic core, however only helices 1, 2, 4 and 5 are involved in native tertiary interactions in the transition state. Helices 3 and 6 interact with the core helices on a later folding time-scale.