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Human Multiple Inositol Polyphosphate Phosphatase as Apoptosis Biomarker
Author(s) -
Hossain Md Zakir,
Kleve Maurice G.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.943.10
Subject(s) - inositol , polyclonal antibodies , biochemistry , phosphatase , microbiology and biotechnology , biology , alkaline phosphatase , apoptosis , antibody , enzyme , chemistry , receptor , immunology
The multiple inositol polyphosphate phosphatase (MINPP) which regulates cellular levels of inositol phosphates, is located on chromosome 10q23 in human cells and is a member of histidine phosphatases family. In this study, we have determined the antigenicity of MINPP. We proposed different possible peptides and designed the experimental methodology to produce MINPP polyclonal antibodies. To further elucidate the physiological role of MINPP we have characterized the human MINPP antibodies using direct western immunoblotting of bacteria (E. coli), yeast (Saccharomyces cerevisae) expressing MINPP. SDS‐PAGE identified 58 KDa protein band using mammalian pancreatic cancer cell lysates (Panc1) and homogenate from rat liver and microsomal fraction of rat liver and kidney after immunoprecipitations with MINPP antibodies. The MINPP enzyme activity was determined by two methods. First, inositol phosphates separation by column chromatography using the microsomal fraction of rat liver and kidney, total cell lysates from rat liver in a scintillation counter using tritiated‐inositol (3H‐inositol). Second, a customized colorimetric method was developed without any radio‐active 3H ‐inositol using 96 wells plate reader. Using different apoptosis inducers, we have seen that MINPP expression increased in programmed cell death in a dose dependent manner. Supported by University of Arkansas at Little Rock.