Regional assignment of the human gene for platelet-type phosphofructokinase (PFKP) to chromosome 10p: novel use of polyspecific rodent antisera to localize human enzyme genes.
Human phosphofructokinase (PFK; EC 2.7.1.11) is under the control of three structural loci which encode muscle-type (M), liver-type (L), and platelet or fibroblast-type (P) subunits; human diploid fibroblasts and leukocytes express all three loci. In order to assign the human PFKP locus to a specific human chromosome, in this study, we have examined ten human X rodent somatic cell hybrids for the expression of human P subunits using a mouse anti-human P subunit-specific antiserum in an active-enzyme-immunoprecipitation technique. In nine of ten hybrids studied, the expression of the PFKP locus segregated concordantly with chromosome 10 and none other, indicating that PFKP is located on chromosome 10; the discordancy rates for all the other chromosomes were 0.2 or greater. In the one discordant hybrid, only the long arm of chromosome 10 was retained and PFKP was not expressed. Human fibroblasts from a patient with duplication of the short arm of chromosome 10 consistently exhibited PFK activity values 180% of normal. These data indicate that human PFKP is located on the short arm of chromosome 10, and that a gene dosage effect is demonstrable in fibroblasts with a duplication of 10p. The use of rodent antihuman antibody combined with immunoprecipitation aided by staphylococci-bearing protein A may find general application in mapping human enzyme genes, when human and rodent gene-products are not distinguishable by other means.
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001792:Blood Platelets / Q000201:enzymology*; D002874:Chromosome Mapping; D002906:Chromosomes, Human, 6-12 and X; D006224:Cricetinae; D003412:Cricetulus; D005347:Fibroblasts / Q000201:enzymology / Q000648:ultrastructure; D006801:Humans; D006822:Hybrid Cells; D007106:Immune Sera; D010732:Phosphofructokinase-1 / Q000235:genetics*; D051381:Rats
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D010732:Phosphofructokinase-1