Summary
A ribonuclease (RNase) was isolated from the urine of a 35-year-old male and purified to electrophoretic homogeneity. The enzyme was tentatively designated RNase 2. A rabbit antibody produced by injection of the purified RNase 2 was able to distinguish RNase 2 from another type of RNase coexisting in body fluids. With this antibody it was possible to detect RNase 2 isozymes in human serum and urine without difficulty using isoelectric focusing or sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by immunoblotting. Both RNase 2 in serum and urine seemed to exist in multiple forms with regard to their molecular masses and pI values. This technique may prove to be useful in genetic and forensic studies of RNase polymorphism.
Zusammenfassung
Eine Ribonuclease (RNase) wurde aus Urin eines gesunden Mannes (35 Jahre) isoliert und bis zur elektrophoretischen Homogenität gereinigt. Das Enzym wurde versuchsweise “RNase 2” genannt. Ein beim Kaninchen hergestellter Antikörper gegen RNase 2 gestattete die Unterscheidung von RNase 2 und einer anderen in Körperflüssigkeiten vorkommenden RNase. Nach Polyacrylamidgel-isoelektrischer Fokussierung oder Natrium Dodecylsulfat-Polyacrylamidgel-Elektrophorese und Immunoblotting mit dem Antikörper war der Nachweis von RNase 2-Isozymen sowohl im Urin als auch in Serum möglich. RNase 2 scheint in verschiedenen Formen in Bezug auf das Molekulargewicht und den isoelektrischen Punkt (pI) im Urin und Serum vorzukommen. Weitere Untersuchungen zum RNase-Polymorphismus stehen noch aus, könnten aber nützliche Anwendungen in der Forensik aufdecken.
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Supported in part by grants from the Japan Brain Foundation and Tokyo Immunopharmacology Institute, and a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan
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Mizuta, K., Yasuda, T., Ikehara, Y. et al. New detection method for ribonuclease 2 (RNase 2) using immunoblotting with specific antibody. Z Rechtsmed 103, 315–322 (1990). https://doi.org/10.1007/BF00204451
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DOI: https://doi.org/10.1007/BF00204451