Calcium removal from Calmodulin
Gabriel Akopian
akopian at oxy.edu
Thu Mar 30 19:42:25 EST 1995
Can anyone, someone, please.....please....send me suggestions on how
to remove calcium from calmodulin. We've been using a chelex-100
column (1cm x 20cm) in all sorts of buffers (ammonium bicarb, sodium
acetate) at various pH levels (from 7.5 to 9.0). At these buffered
pH ranges we've only managed to remove 3 out of the 4 Ca2+ that bind
to calmodulin. When we don't use any buffers, (pH of chelex 10 to 12)
we can remove all four Ca2+. The only problem is, that at this pH
range, the protein begins to denature (based on UV spec). My guess is
that at this pH the Ca2+ binding site that refuses to let go of the
Ca2+ becomes exposed (because of denaturation) and thus can readily
give up the last Ca2+ to the chelex. Does anyone have any ideas....
Also, all metal ion determinations are done by Atomic Absorbance
measurements.
More information about the Methods
mailing list