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Literature summary for 3.1.3.1 extracted from

  • Numa, N.; Ishida, Y.; Nasu, M.; Sohda, M.; Misumi, Y.; Noda, T.; Oda, K.
    Molecular basis of perinatal hypophosphatasia with tissue-nonspecific alkaline phosphatase bearing a conservative replacement of valine by alanine at position 406. Structural importance of the crown domain (2008), FEBS J., 275, 2727-2737.
    View publication on PubMed

Application

Application Comment Organism
medicine this particular V406A mutation highlights the structural importance of the crown domain with respect to the catalytic function of TNSALP Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expressed in transiently transfected COS-1 cells and in stable CHO-K1 Tet-On cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
V406A mutant protein shows a markedly reduced alkaline phosphatase activity. Mutant is conveyed to the Golgi apparatus and incorporated into a cold detergent insoluble fraction (raft) at a rate similar to that of the wild-type. Mutant shows increased susceptibility to digestion by trypsin or proteinase K. When purified as a glycosylphosphatidylinositol-anchorless soluble form, mutant protein exhibits a remarkably lower Kcat /Km value compared with that of the wild-type TNSALP Homo sapiens
V406F replacement with phenylalanine results in a low enzyme activity, even though TNSALP (V406F) is processed to the 80 kDa mature form similarly to TNSALP (V406A) and appears on the cell surface like the wild-type protein Homo sapiens
V406I similar to wild-type, leucine and isoleucine can be successfully substituted for the valine residue Homo sapiens
V406L similar to wild-type, leucine and isoleucine can be successfully substituted for the valine residue Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.09
-
4-nitrophenyl phosphate pH 10.5, mutant V406A Homo sapiens
0.21
-
4-nitrophenyl phosphate pH 10.5, wild-type Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
immunoblotting, immature form bearing high mannose-type N-linked oligosaccharides Homo sapiens
80000
-
immunoblotting, mature form bearing complex-type oligosaccharides Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
Homo sapiens 4-nitrophenol + phosphate
-
?

Subunits

Subunits Comment Organism
homodimer sucrose density gradient centrifugation, wild-type and V406A form a homodimer Homo sapiens

Synonyms

Synonyms Comment Organism
tissue-nonspecific alkaline phosphatase
-
Homo sapiens
TNSALP
-
Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
34
-
4-nitrophenyl phosphate pH 10.5, mutant V406A Homo sapiens
971
-
4-nitrophenyl phosphate pH 10.5, wild-type Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10.5
-
assay at Homo sapiens