The occurrence of collagen in the cockroach Leucophaea maderae has been demonstrated by electron optical and biochemical techniques. Electron micrographs of tissues of this and a related species ( Blaberus craniifer ) are presented and they show that collagenous-type fibers occur in the stroma of nonneural as well as neural organs of these insects. Hydroxyproline and hydroxylysine, amino acids considered to be "markers" for collagen, have been shown to be present in proteins extracted from material rich in neuroglandular tissue (corpus cardiacum plus corpus allatum). Trimmed carcasses of Leucophaea maderae have been shown to contain a protein or proteins soluble in hot trichloroacetic acid, with compositional characteristics similar to those of collagens in general, including diagnostic proportions of glycine, proline, hydroxyproline, and hydroxylysine. This collagen is not soluble in dilute acetic acid or in concentrated solutions of guanidinium chloride. It is measurably digested by bacterial collagenase.
1. Purified citrate-extracted ichthyocol obtained from carp swim bladders has been further characterized with respect to its content of certain amino acids and carbohydrate substances. 2. The degree of solubilization or dispersion of ichthyocol by solutions of certain salts maintained in the range of neutral pH and at a temperature of 0–2°C. has been determined. 3. While a number of salts of monovalent cations had no significant solubilizing effects on ichthyocol, ammonium chloride in a concentration of 1 M did cause solution of the protein. 4. Sodium thiosulfate in a range of concentrations caused the solubilization of ichthyocol but was most effective in an intermediate concentration of 0.25 M . 5. Several salts of divalent cations, in particular the chlorides of calcium, magnesium, and barium, and magnesium thiosulfate in concentrations ranging from 0.3 to 1 M caused the immediate and complete solubilization of the ichthyocol. 6. Solutions of ichthyocol in calcium chloride, magnesium chloride, and sodium thiosulfate buffered or adjusted to pH 7.0, were studied with respect to intrinsic viscosity of the protein, optical rotation, ultracentrifugal sedimentation, and reconstitution into fibers. It was found in each case that the original characteristics of the collagen, as determined previously in acid solution, were maintained when the protein was dissolved in salt solutions of neutral pH. No evidence of denaturation or gelatinization could be found when ichthyocol was solubilized under the stated conditions. 7. Collagen in neutral solution with sodium thiosulfate, calcium chloride, or magnesium chloride was not attacked by trypsin as determined viscometrically at 20.0°C., but was rapidly degraded by a purified bacterial collagenase.