Size and shape of the repetitive domain of high molecular weight wheat gluten proteins. II. Hydrodynamic studies.
This study describes the hydrodynamic properties of the repetitive domain of high molecular weight (HMW) wheat proteins, which complement the small-angle scattering (SANS) experiments performed in the first paper of this series. The sedimentation coefficients, s(0), and diffusion coefficients, D(0), were obtained from the homologous HMW proteins dB1 and dB4 that were cloned from the gluten protein HMW Dx5, and expressed in Escherichia coli. Monodisperse conditions for accurate determination of s(0) and D(0), were obtained by screening a series of buffers using dynamic light scattering. For the first time, hydrodynamic parameters were obtained on monodisperse samples that enabled the determination of the monomeric size and shape. The hydrodynamic values determined on dB1 and dB4 were used to test the worm-like chain (WLC) model that was proposed in the SANS studies. The successful matching of two separately obtained hydrodynamic parameters of dB1 and dB4 using the WLC model provides further evidence for the WLC model. The small discrepancy between the hydrodynamic and scattering data, possibly coming from the excluded volume effect, was compensated by a solvation layer of 1-2 water molecules thick around the protein in the WLC model. The solvation of the central domain is much higher than those of the terminal domains of the HMW subunits. This difference emphasizes the dual role of HMW wheat gluten proteins in water-binding and aggregation.
van Swieten
Eric
E
University of Groningen, Department of Biochemistry, and Groningen Biomolecular Sciences and Biotechnology Institute, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
Friesen
Robert R
RR
de Kruif
Cees G
CG
Robillard
George T
GT
D001704:Biopolymers; D002021:Buffers; D017276:Friction; D005983:Glutens / Q000737:chemistry*; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D012542:Scattering, Radiation; D014908:Triticum / Q000737:chemistry*; D014461:Ultracentrifugation; D014867:Water / Q000737:chemistry*
D001704:Biopolymers; D002021:Buffers; D014867:Water; D005983:Glutens