Radioimmune determination of hypoxanthine phosphoribosyltransferase crossreacting material in erythrocytes of Lesch--Nyhan patients

Journal Article · Wed Oct 01 04:00:00 EDT 1975 · Proc. Natl. Acad. Sci. U.S.A., v. 72, no. 10, pp. 4147-4150

DOI:https://doi.org/10.1073/pnas.72.10.4147· OSTI ID:4142380

Ghangas, G S; Milman, G

We have developed a sensitive radioimmunoassay capable of detecting and quantitating 20 ng of hypoxanthine phosphoribosyltransferase (EC 2.4.2.8; IMP: pyrophosphate phosphoribosyltransferase) protein. For this assay, hypoxanthine phosphoribosyltransferase from human erythrocytes was iodinated with $sup 125$I under mild conditions using hydrogen peroxide and lactoperoxidase attached to Sepharose-4B. Antisera prepared against homogeneous human hypoxanthine phosphoribosyltransferase precipitates the iodinated enzymes as effectively as the unlabeled enzyme. The radioimmunoassay has been used to look for hypoxanthine phosphoribosyltransferase crossreacting material in hemolysates from sixteen different patients with a marked genetic deficiency of this enzyme characteristic of the Lesch--Nyhan syndrome. Fifteen hemolysates contained no detectable crossreacting material. One hemolysate contained a normal amount of crossreacting material. Hypoxanthine phosphoribosyltransferase from this patient has been shown to be a K/sub m/ mutant enzyme. (auth)

• Research Organization: Univ. of California, Berkeley
• Sponsoring Organization: USDOE
• NSA Number: NSA-33-012546
• OSTI ID: 4142380
• Journal Information: Proc. Natl. Acad. Sci. U.S.A., v. 72, no. 10, pp. 4147-4150, Journal Name: Proc. Natl. Acad. Sci. U.S.A., v. 72, no. 10, pp. 4147-4150; ISSN PNASA
• Country of Publication: United States
• Language: English

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