Calcium and the assays of human plasma clotting factor XIII.
1. A continuous fluorimetric assay for blood-clotting Factor XIII based on the incorporation of dansylcadaverine into casein was investigated. 2. Hammarsten casein was fractionated to yield the beta-casein, which was dephosphorylated and acetylated to give a substrate which itself did not bind calcium and which was not cross-linked by the activated Factor. 3. The modified beta-casein was used as substrate for a continuous fluorimetric assay and as substrate for the incorporation of radioactive glycine ethylester. 4. A linked fluorimetric assay for the zymogen is described. 5. The K(m) for calcium was redetermined and at 0.2mm was in the physiological range and much lower than the values reported by others using substrates which interact with calcium. 6. The K(m) for casein is about 14mum.
D000107:Acetylation; D002118:Calcium; D002250:Carbon Radioisotopes; D002364:Caseins; D002848:Chromatography, DEAE-Cellulose; D003619:Dansyl Compounds; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors; D005176:Factor XIII / Q000032:analysis*; D005998:Glycine; D006801:Humans; D007700:Kinetics; D012967:Sodium Dodecyl Sulfate; D013050:Spectrometry, Fluorescence
D002250:Carbon Radioisotopes; D002364:Caseins; D003619:Dansyl Compounds; D004792:Enzyme Precursors; D012967:Sodium Dodecyl Sulfate; D005176:Factor XIII; D002118:Calcium; D005998:Glycine