Purification, Peptide Seaquencing, Chararcterisation and Study of antiproliferative activity of...
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![Page 1: Purification, Peptide Seaquencing, Chararcterisation and Study of antiproliferative activity of Cicer arietinum L. Protein phosphatase 2A on Chronic Myelogenous Leukemia Cell lines.](https://reader035.fdocuments.us/reader035/viewer/2022081210/55c28536bb61eba6438b45f4/html5/thumbnails/1.jpg)
Purification , Peptide Sequencing ,Characterisation and Study of
Antiproliferative activity of Cicer arietinum L. Protein
Phosphatase 2A on Chronic Myelogenous Leukemia K562 Cell line:
Ellucidation of Mechanism of Development of Chronic
Myelogenous Leukemia in Human Cell line.
Antik Kiron Bose Ph.D 1
1 Senior Scientist and Project Advisor, Human Biology Division, Fred Hutchinson Cancer Research
Center,Seattle, Washington.
Protein Phosphatase 2A has been purified from 72 hrs germinating
seeds of Cicer arietinum L. by Toyopearl DEAE -650 anion exchanger,
Phenyl-sepharose CL-4B ,Microcystin –aminoethanethiol- sepharose
chromatography ,PD-10 desalting column and Vydac C-18 Reverse phase
HPLC chromatography with 4186.67 folds of purification ,3% recovery
and final specific activity of 209.334 U/ �g protein /min .The molar
masses of catalytic subunit (38 KD) and regulatory subunits (65 and 75
KD) were determined by 6% SDS-PAGE .38 KD catalytic subunit was
identified by Western Blotting with anti IgG2 monoclonal Ab against
human PP2A catalytic subunit.Number of SH groups was found to be 11
in PP2A/c and its full peptide sequencing gave a 306 aa protein which
was deposited in Protein Public database Uniprot with Acc no. Q07098
.Human PP2A was found to be inhibited in human CML cell line K562 due
to Bcr:abl induced formation of binary complex between human PP2A/c
and hnRNP A2 and ternary complex between human PP2A holoenzyme,hn
RNP A2 and SET protein .In K562 mutant OA200 cell line lacking human
PP2A ; chick pea PP2A was found to induce apoptosis of CML cells as it
is not inhibited by hn RNP A2 or SET proteins in vivo or in
vitro.Chick pea PP2A further induced nuclear localization of
![Page 2: Purification, Peptide Seaquencing, Chararcterisation and Study of antiproliferative activity of Cicer arietinum L. Protein phosphatase 2A on Chronic Myelogenous Leukemia Cell lines.](https://reader035.fdocuments.us/reader035/viewer/2022081210/55c28536bb61eba6438b45f4/html5/thumbnails/2.jpg)
SET-NUP214 complex and degradation of bcr:abl fusion proteins .RNA
binding domains of hn RNP A2 was sufficient to inhibit human PP2A .
Chick pea PP2A associated with human PR72 and induced
dephosphorylation of SV 40 T antigen. pH and temperature optima of
chick pea PP2A were 8.0 and 25 �c respectively and Km and V max were
87.713 mM , 166.67 U/min for holoenzyme and 64.10 mM and 208.33 U/min
for PP2A respectively.