Stereochemistry of collagen.

This review article, based on a lecture delivered in Madras in 1985, is an account of the author's experience in the working out of the molecular structure and conformation of the collagen triple-helix over the years 1952-78. It starts with the first proposal of the correct triple-helix in 1954, but with three residues per turn, which was later refined in 1955 into a coiled-coil structure with approximately 3.3 residues per turn. The structure readily fitted proline and hydroxyproline residues and required glycine as every third residue in each of the three chains. The controversy regarding the number of hydrogen bonds per tripeptide could not be resolved by X-ray diffraction or energy minimization, but physicochemical data, obtained in other laboratories during 1961-65, strongly pointed to two hydrogen bonds, as suggested by the author. However, it was felt that the structure with one straight NH...O bond was better. A reconciliation of the two was obtained in Chicago in 1968, by showing that the second hydrogen bond is via a water molecule, which makes it weaker, as found in the physicochemical studies mentioned above. This water molecule was also shown, in 1973, to take part in further cross-linking hydrogen bonds with the OH group of hydroxyproline, which occurred always in the location previous to glycine, and is at the right distance from the water. Thus, almost all features of the primary structure, X-ray pattern, optical and hydrodynamic data, and the role of hydroxyproline in stabilising the triple helical structure, have been satisfactorily accounted for. These also lead to a confirmation of Pauling's theory that vitamin C improves immunity to diseases, as explained in the last section.

A theoretical study of the structures of (Gly-Pro-Leu)n and (Gly-Leu-Pro)n.

Theoretical conformation studies have been carried out for the polytripeptides (Gly-Pro-Leu)n and (Gly-Leu-Pro)n and the Fourier transforms of the structures have been calculated. X-ray powder patterns of these polymers had indicated that both these polymers take up coiled-coil triple-helical structures, but in the case of (Gly-Pro-Leu)n it was not clear whether the triple helix is formed by three parallel polypeptide chains or by a single chain folding back on itself (Scatturin et al 1975). Our studies show that both the polytripeptides can take up stereochemically satisfactory triple-helical structures with three parallel chains. There is also very good agreement between the calculated intensity distribution and that of the observed X-ray pattern, in each case.

Three-dimensional reconstruction from radiographs and electron micrographs: application of convolutions instead of Fourier transforms.

A new technique is proposed for the mathematical process of reconstruction of a three-dimensional object from its transmission shadowgraphs; it uses convolutions with functions defined in the real space of the object, without using Fourier transforms. The object is rotated about an axis at right angles to the direction of a parallel beam of radiation, and sections of it normal to the axis are reconstructed from data obtained by scanning the corresponding linear strips in the shadowgraphs at different angular settings. Since the formulae in the convolution method involve only summations over one variable at a time, while a two-dimensional reconstruction with the Fourier transform technique requires double summations, the convolution method is much faster (typically by a factor of 30); the relative increase in speed is larger where greater resolution is required. Tests of the convolution method with computer-simulated shadowgraphs show that it is also more accurate than the Fourier transform method. It has good potentialities for application in electron microscopy and x-radiography. A new method of reconstructing helical structures by this technique is also suggested.