Negative staining and genesis of D-periodicity in native collagen fibrils

An investigation was carried out on the mechanism which gives rise to the banding exhibited by collagen fibrils after negative staining. The negative staining (phosphotungstic acid) band patterns of native collagen fibrils (type I), isolated from calf reticular dermis, were compared with computer-drawn band patterns. The stimulations were based on the primary structure of bovine type I collagen, the "quarter stagger" molecular packing and different conformations of telopeptides. The results suggest that in negative staining, the stain exclusion effect depends on both spatial factors and water repelling factors being the "bulkiness" (molecular volume/length ratio) as well as the hydrophobicity of the amino acids of alpha 1 (I) and alpha 2 (I) chains directly involved. No final conclusion could be drawn about the contribution of positive staining to negative staining. Improvements in the simulations were achieved when the telopeptides were shaped according to particular conformational models.