Self-association of streptococcus pyogenes collagen-like constructs into higher order structures.

A number of bacterial collagen-like proteins with Gly as every third residue and a high Pro content have been observed to form stable triple-helical structures despite the absence of hydroxyproline (Hyp). Here, the high yield cold-shock expression system is used to obtain purified recombinant collagen-like protein (V-CL) from Streptococcus pyogenes containing an N-terminal globular domain V followed by the collagen triple-helix domain CL and the modified construct with two tandem collagen domains V-CL-CL. Both constructs and their isolated collagenous domains form stable triple-helices characterized by very sharp thermal transitions at 35-37 degrees C and by high values of calorimetric enthalpy. Procedures for the formation of collagen SLS crystallites lead to parallel arrays of in register V-CL-CL molecules, as well as centrosymmetric arrays of dimers joined at their globular domains. At neutral pH and high concentrations, the bacterial constructs all show a tendency towards aggregation. The isolated collagen domains, CL and CL-CL, form units of diameter 4-5 nm which bundle together and twist to make larger fibrillar structures. Thus, although this S. pyogenes collagen-like protein is a cell surface protein with no indication of participation in higher order structure, the triple-helix domain has the potential of forming fibrillar structures even in the absence of hydroxyproline. The formation of fibrils suggests bacterial collagen proteins may be useful for biomaterials and tissue engineering applications.