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

↓ Direkt till sidans innehåll
↓ Direkt till sidans sekundära innehåll (sidomenyn)

Search: onr:"swepub:oai:DiVA.org:umu-20842" > Structure, genomic ...

1 of 1
Previous record
Next record
To hitlist

Details
MARC

Wennberg, Charlotte (author)

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

Article/chapterEnglish2002

Publisher, publication year, extent ...

2002-02-13
Hindawi Limited,2002
printrdacarrier

Numbers

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

Supplementary language notes

Language:English
Summary in:English

Part of subdatabase

SwePubSwePub

Classification

Subject category:ref swepub-contenttype
Subject category:art swepub-publicationtype

Notes

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.

Subject headings and genre

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

Added entries (persons, corporate bodies, meetings, titles ...)

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

Related titles

In:Human Mutation: Hindawi Limited19:3, s. 258-2671059-77941098-1004

Internet link

Find in a library

Human Mutation (Search for host publication in LIBRIS)

To the university's database

1 of 1
Previous record
Next record
To hitlist

Find more in SwePub

By the author/editor: Wennberg, Charlo ...; Kozlenkov, Alexe ...; Mauro, Sonia Di; Fröhlander, Nils; Beckman, Lars; Hoylaerts, Marc ...; show more...; Millán, José Lui ...; show less...

Articles in the publication: Human Mutation

By the university: Umeå University

Search outside SwePub

Extend your search to:: Google; Google Book Search; Google Scholar

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.

LIBRIS.kb.se