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WFRF:(Millán José Luis)
 

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  • Wennberg, Charlotte (författare)

Structure, genomic DNA typing and kinetic characterization of the D allozyme of placental alkaline phosphatase

  • Artikel/kapitelEngelska2002

Förlag, utgivningsår, omfång ...

  • 2002-02-13
  • Hindawi Limited,2002
  • printrdacarrier

Nummerbeteckningar

  • LIBRIS-ID:oai:DiVA.org:umu-20842
  • https://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-20842URI
  • https://doi.org/10.1002/humu.10052DOI

Kompletterande språkuppgifter

  • Språk:engelska
  • Sammanfattning på:engelska

Ingår i deldatabas

Klassifikation

  • Ämneskategori:ref swepub-contenttype
  • Ämneskategori:art swepub-publicationtype

Anmärkningar

  • The 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 wild-type Chinese hamster ovary cells. We determined the kcat and Km, 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.

Ämnesord och genrebeteckningar

  • alkaline phosphatase
  • placental; PLAP; ALPP; ALPL; ALPPL2; ALPI; isozyme; negative selection; spontaneous abortion; gene therapy; genetic disease; placental function; SNuPE

Biuppslag (personer, institutioner, konferenser, titlar ...)

  • Kozlenkov, AlexeyUmeå universitet,Institutionen för medicinsk biovetenskap (författare)
  • Mauro, Sonia Di (författare)
  • Fröhlander, Nils (författare)
  • Beckman, Lars (författare)
  • Hoylaerts, Marc F. (författare)
  • Millán, José Luis (författare)
  • Umeå universitetInstitutionen för medicinsk biovetenskap (creator_code:org_t)

Sammanhörande titlar

  • Ingår i:Human Mutation: Hindawi Limited19:3, s. 258-2671059-77941098-1004

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