A review of multi-domain and flexible molecular chaperones studies by small-angle X-ray scattering

Abstract

Intrinsic flexibility is closely related to protein function, and a plethora of important regulatory proteins have been found to be flexible, multi-domain or even intrinsically disordered. On the one hand, understanding such systems depends on how these proteins behave in solution. On the other, small-angle X-ray scattering (SAXS) is a technique that fulfills the requirements to study protein structure and dynamics relatively quickly with few experimental limitations. Molecular chaperones from Hsp70 and Hsp90 families are multi-domain proteins containing flexible and/or disordered regions that play central roles in cellular proteostasis. Here, we review the structure and function of these proteins by SAXS. Our general approach includes the use of SAXS data to determine size and shape parameters, as well as protein shape reconstruction and their validation by using accessory biophysical tools. Some remarkable examples are presented that exemplify the potential of the SAXS technique. Protein structure can be determined in solution even at limiting protein concentrations (for example, human mortalin, a mitochondrial Hsp70 chaperone). The protein organization, flexibility and function (for example, the J-protein co-chaperones), oligomeric status, domain organization, and flexibility (for the Hsp90 chaperone and the Hip and Hep1 co-chaperones) may also be determined. Lastly, the shape, structural conservation, and protein dynamics (for the Hsp90 chaperone and both p23 and Aha1 co-chaperones) may be studied by SAXS. We believe this review will enhance the application of the SAXS technique to the study of the molecular chaperones.

Keywords: Molecular chaperones; Mortalin Hsp70; Multi-domain proteins; Protein dynamics; SAXS.

Fig. 1

Domain architecture of Hsp70 and Hsp90 molecular chaperones families and some of their co-chaperones that form the focus in this review. The abbreviated names of the domains are placed above the domain (see text for identification). The size of the domains and regions corresponds to their peptide lengths. Protein domains are described in the text. Zigzag lines represent flexible regions or intrinsic unfolded sequences

Fig. 2

Small-angle X-ray scattering experimental setup