The mean residue ellipticity, [theta], at any wavelength, lambda, of a protein in aqueous solution is expressed as [theta]lambda = fH[theta]H infinity(1-k/n) + f beta[theta]beta + ft[theta]t + fR[theta]R with two constraints: 1 > or = fj > or = 0 and sigma fj = 1. The subscripts H, beta, t, and R refer to the helix, beta-form, beta-turn, and unordered form. The fractions, fj's, of 15 proteins are based on X-ray crystallography, ft refers to the net beta-turn after cancelling those residues having dihedral angles of opposite sign. The [theta]H infinity of an infinite helix and its chain-length dependence factor, k, were computed from the myoglobin data (Chen et al., 1974, Biochemistry 13, 3350). The average number of residues per helical segment, n, for 15 proteins was about 10, which can be used for proteins of unknown structure. The reference spectra of other three structural elements are computed by a least-squares method. Once the reference spectra are chosen, the same equation above can be used to estimate the fractions of the secondary structure of a protein from its CD data points between 190 and 240 nm at 1-nm intervals. The computed helical content is usually good to excellent (concanavalin A is a notable exception). Inclusion of the beta-turn in the analysis improves the correlation for the estimates of the beta-form, but the computed beta t values are not significantly correlated with the X-ray results. Matrix formulation proves the equivalence of the least-squares method and the integral curve-fitting.