SummaryAn indirect spectrophotometric assay for extrinsic plasminogen activator has been devised, which is based on the parabolic assay of Drapier et al. (5). The system contains activator, plasminogen, the synthetic plasmin substrate H-D-Val-Leu-Lys-pNA (S-2251, Kabi) and a mixture of soluble fibrinogen fragments prepared by treatment of fibrinogen with cyanogen bromide. The addition of these fibrinogen fragments considerably enhances the sensitivity and specificity of the method owing to specific stimulation of the plasminogen activation by extrinsic plasminogen activator.The assay conditions were optimized and the application for extrinsic plasminogen activator measurements in plasma euglobulin fractions is demonstrated.

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The soluble potato proteins are mainly composed of patatin and protease inhibitors. Using DSC and both far-UV and near-UV CD spectroscopy, it was shown that potato proteins unfold between 55 and 75 degrees C. Increasing the ionic strength from 15 to 200 mM generally caused an increase in denaturation temperature. It was concluded that either the dimeric protein patatin unfolds in its monomeric state or its monomers are loosely associated and unfold independently. Thermal unfolding of the protease inhibitors was correlated with a decrease in protease inhibitor activities and resulted in an ionic strength dependent loss of protein solubility. Potato proteins were soluble at neutral and strongly acidic pH values. The tertiary structure of patatin was irreversibly altered by precipitation at pH 5. At mildly acidic pH the overall potato protein solubility was dependent on ionic strength and the presence of unfolded patatin.

The effects of pH and various additives on the precipitation and (re)solubility at pH 7 of potato proteins from industrial potato fruit juice (PFJ) were studied. The use of various strong and weak acids did not result in differences in protein precipitation, which on average occurred to a maximum of 60% of total protein at pH 3. Weak acids did, however, result in precipitates with a higher resolubility at pH 7 compared with strong acids. At pH 5, addition of FeCl 3 or ZnCl 2 increased both precipitation and resolubility compared with the situation without additives. The largest increases in both precipitation and resolubility were achieved using organic solvents at pH 5 as an additive, resulting in a maximum precipitation and maximum resolubility at pH 7 of 91 and 83% of total protein respectively. The results described in this study lead to the hypothesis that precipitation and resolubilisation of potato proteins from PFJ are not determined by their isoelectric pH but by their interactions with low-molecular-weight components.