Expression of a heterodimeric (placental-intestinal) hybrid alkaline phosphatase in KB cells
| Autor: | Torgny Stigbrand, Tetsuo Adachi, Kyozo Hayashi, Kentaro Asai, Yukio Mori, Kazuyuki Hirano, Hirohiko Kodama |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Meconium
Placenta Phosphatase Molecular Sequence Data Biophysics Gene Expression Biology Biochemistry Isozyme KB Cells Amidohydrolases chemistry.chemical_compound Structural Biology Genetics Humans Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Amino Acid Sequence Cyanogen Bromide Peptide sequence chemistry.chemical_classification Gel electrophoresis Tunicamycin Antibodies Monoclonal Alkaline Phosphatase Molecular biology Peptide Fragments Amino acid Intestines Isoenzymes Molecular Weight Placental alkaline phosphatase chemistry Alkaline phosphatase Cyanogen bromide |
| Zdroj: | Biochimica et biophysica acta. 1218(2) |
| ISSN: | 0006-3002 |
| Popis: | A hybrid heterodimeric alkaline phosphatase expressed in KB cells, consisting of placental and intestinal (fetal) subunits, was purified by use of two different immunoaffinity columns using the monoclonal antibodies 2HIMS-1 and HPMS-1. The closely related subunits were found to yield a dimeric active enzyme glycosylated as the mature heterodimeric forms. This enzyme displays intermediate properties to the placental and intestinal (fetal) isozymes with regard to heat stability, inhibition patterns with amino acids and amino acid derivatives, as well as reactivity with monoclonal antibodies specific for human alkaline phosphatase isozymes. Peptide fragments obtained from the hybrid enzyme after cyanogen bromide cleavage belong to either the placental or intestinal (meconial) isozyme as evaluated by SDS polyacrylamid gel electrophoresis, and the N-terminal amino acid sequences, corresponding to the placental and intestinal subunits, can be identified in the peptide fragments. By N-glycanase digestion or tunicamycin treatment, the molecular mass of the subunits was reduced to 62 kDa compared to 69 kDa for the native ones. The results confirm that some cell lines can synthesize hybrid alkaline phosphatases. |
| Databáze: | OpenAIRE |
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