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Literature summary for 3.1.3.1 extracted from

  • Zhang, F.; Murhammer, D.W.; Linhardt, R.J.
    Enzyme kinetics and glycan structural characterization of secreted alkaline phosphatase prepared using the baculovirus expression vector system (2002), Appl. Biochem. Biotechnol., 101, 197-210.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Spodoptera frugiperda Sf9 and Trichoplusia ni BTI-Tn-5B1-4 insect cells via infection with recombinant Autographa california multiple nuclear polyhedrovirus expressing SEAP under control of the polyhedrin promotor, the recombinant enzyme expressed in the 2 different insect cell lines show differing N-glycan structures Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.59
-
4-nitrophenyl phosphate pH 9.8, recombinant enzyme expressed in Tn-5B1-4 cells Homo sapiens
6.76
-
4-nitrophenyl phosphate pH 9.8, recombinant enzyme expressed in Sf-9 cells Homo sapiens
49.7
-
4-nitrophenyl phosphate pH 9.8, native placental enzyme Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
64000
-
1 * 64000, recombinant enzyme, SDS-PAGE, 1 * 66200, native enzyme, SDS-PAGE Homo sapiens
65000
-
about, recombinant enzyme, gel filtration Homo sapiens
66200
-
native placental enzyme, gel filtration Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
secreted alkaline phosphatase SEAP containing a single N-glycan chain, a model protein for investigation of glycosylation pattern
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein secreted alkaline phosphatase SEAP containing a single N-glycan chain, a model protein for investigation of glycosylation patterns, recombinant enzymes expressed in insect cells shows differing patterns: Sf-9 cell derived enzyme has Mna4GlcNAc2 and FucMan4GlcAc2 as major N-glycans, while the Tn-5B1-4 cell derived enzyme has Man2GlcAc2 as major N-glycan, glycan structures overview Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from expression in insect cells Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
a phosphate monoester + H2O = an alcohol + phosphate Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
placenta placental SEAP Homo sapiens
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
629
-
purified recombinant enzyme expressed in Sf-9 cells Homo sapiens
738
-
purified native placental enzyme Homo sapiens
850
-
purified recombinant enzyme expressed in Tn-5B1-4 cells 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
monomer 1 * 64000, recombinant enzyme, SDS-PAGE, 1 * 66200, native enzyme, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
SEAP
-
Homo sapiens
secreted alkaline phosphatase
-
Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.8
-
assay at Homo sapiens