Structure, genomic DNA typing, and kinetic characterization of the D allozyme of placental alkaline phosphatase (PLAP/ALPP)

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorWennberg, C.
dc.creatorKozlenkov, A.
dc.creatorDi Mauro, S.
dc.creatorFrohlander, N.
dc.creatorBeckman, L.
dc.creatorHoylaerts, M. F.
dc.creatorMillan, J. L.
dc.date2014-05-20T15:24:07Z
dc.date2016-10-25T17:58:13Z
dc.date2014-05-20T15:24:07Z
dc.date2016-10-25T17:58:13Z
dc.date2002-01-01
dc.date.accessioned2017-04-05T23:45:30Z
dc.date.available2017-04-05T23:45:30Z
dc.identifierHuman Mutation. New York: Wiley-liss, v. 19, n. 3, p. 258-267, 2002.
dc.identifier1059-7794
dc.identifierhttp://hdl.handle.net/11449/34767
dc.identifierhttp://acervodigital.unesp.br/handle/11449/34767
dc.identifier10.1002/humu.10052
dc.identifierWOS:000174215500008
dc.identifierhttp://dx.doi.org/10.1002/humu.10052
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/878626
dc.descriptionThe D allozyme of placental alkaline phosphatase (PLAP) displays enzymatic properties at variance with those of the common PLAP allozymes. We have deduced the amino acid sequence of the PLAP D allele by PCR cloning of its gene, ALPP Two coding substitutions were found in comparison With the cDNA of the common PLAP F allele, i.e., 692C>G and 1352A>G, which translate into a P209R and E429G substitution. A single nucleotide primer extension (SNuPE) assay was developed using PCR primers that enable the amplification of a 1.9 kb PLAP fragment. Extension primers were then used on this PCR fragment to detect the 692C>G and 1352A>G substitution. The SNuPE assay on these two nucleotide substitutions enabled us to distinguish the PLAP F and D alleles from the PLAP S/I alleles. Functional studies on the D allozyme were made possible by constructing and expressing a PLAP D cDNA, i.e., [Arg209, Gly429] PLAP, into wildtype Chinese hamster ovary cells. We determined the k(cat) and K-m, of the PLAP S, F. and D allozymes using the non,physiological substrate p-nitrophenylphosphate at an optimal pH (9.8) as well as two physiological substrates, i.e., pyridoxal-5'-phosphate and inorganic pyrophosphate at physiological pH (7.5). We found that the biochemical properties of the D allozyme of PLAP are significantly different from those of the common PLAP allozymes. These biochemical findings suggest that a suboptimal enzymatic function by the PLAP D allozyme may be the basis for the apparent negative selective pressure of the PLAP D allele. The development of the SNuPE assay will enable us to test the hypothesis that the PLAP D allele is subjected to intrauterine selection by examining genomic DNA from statistically informative population samples. Hum Mutat 19:258-267, 2002. (C) 2002 Wiley-Liss, Inc.
dc.languageeng
dc.publisherWiley-Blackwell
dc.relationHuman Mutation
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectalkaline phosphatase
dc.subjectplacental
dc.subjectPLAP
dc.subjectALPP
dc.subjectALPL
dc.subjectALPPL2
dc.subjectALPI
dc.subjectisozyme
dc.subjectnegative selection
dc.subjectspontaneous abortion
dc.subjectgene therapy
dc.subjectgenetic disease
dc.subjectplacental function
dc.subjectSNuPE
dc.titleStructure, genomic DNA typing, and kinetic characterization of the D allozyme of placental alkaline phosphatase (PLAP/ALPP)
dc.typeOtro


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