Asp361Val Mutant of Alkaline Phosphatase Found in Patients with Dominantly Inherited Hypophosphatasia Inhibits the Activity of the Wild-Type Enzyme

• DOI: 10.1210/jcem.85.2.6373
• Publication Date: 2014-01-08T17:05:11Z
• Authors: Hermann L. Müller; Miwa Yamazaki; Toshimi Michigami; Takashi Kageyama; Eckhard Schönau; Peter Schneider; Keiichi Ozono
• Keywords: Adult; Male; 0301 basic medicine; DNA, Complementary; DNA Mutational Analysis; Gene Expression; Hypophosphatasia; Alkaline Phosphatase; Pedigree; Isoenzymes; 03 medical and health sciences; Mutation; Humans; Female; Amino Acid Sequence; Child; Genes, Dominant

Hypophosphatasia is characterized by the hypomineralization of bone associated with the mutation of the tissue-nonspecific alkaline phosphatase (TNSALP) gene. Although the disease is usually autosomal recessive, an autosomal dominant form is also recognized. Approximately 50 mutations have been found in the TNSALP gene in patients with hypophosphatasia. However, the mutations identified to date do not seem to account for the dominantly inherited form of the disease. We have examined a German family in which the father and all 4 children were affected with hypophosphatasia, whereas the mother was healthy. The affected members of this family showed premature loss of deciduous teeth at or shortly before 2 yr of age and low levels of serum ALP with elevated levels of urinary phosphoethanolamine. DNA analysis by direct sequencing revealed a heterozygous missense mutation that caused the conversion of amino acid Asp to Val at position 361 (D361V) in the patients. Another substitution was detected in exon 12 (Val to Ala conversion at codon 505: V505A) in 1 allele of the mother and 3 children, indicating no association of the substitution with the disease. Reconstruction experiments demonstrated that the D361V mutant protein lost its enzymatic activity and that it inhibited the function of wild-type enzyme when coexpressed in COS-7 cells. On the other hand, the V505A mutant exhibited enzymatic activities equal to those of the wild-type ALP. It is likely that the mutant D361V protein forms dimers with the wild-type protein, and the protein-protein interaction contributes to the dominant effect of the mutant D361V. The mutation that causes D361V is the first one proven to be associated with the dominant form of hypophosphatasia.