Dear George,
For one of the ligand the kd was 50 micromole, the
protein concentration used in MST was 20nM and at
250micromolar ligand concentration, it was ~95% bound.
Thank you
Regards
Kavya
> Almost means 88%? 93% ? 95%? or 99%?
>
> That why it is much easier when Kd is known because we do not need all the
> above detail information just one number (Kd).
> Give me also the protein concentration of protein during bioassay.
>
> G.
>
> -----Original Message-----
> From: [log in to unmask] [mailto:[log in to unmask]]
> Sent: Wednesday, February 10, 2016 1:17 PM
> To: George Kontopidis
> Cc: [log in to unmask]
> Subject: Re: [ccp4bb] Protein:ligand ratio for co-crystallization
>
> Dear George,
>
> Sorry for the late reply. 1:500 the MST curve almost reaches the
> saturation.
>
> Thank you
> Regards
> Kavya
>
>> Dear Kavya,
>>
>> The biophysical techniques like MST shows a binding of 20%, 40% or
>> 99% (protein:ligand)?
>> when molar ratio of protein:ligand is greater than 1:500 we need this
>> number to give you an answer
>>
>> All depend of the Ka or Kd (1/Ka). That why you have obtained ligand
>> bound structure even with molar ratio of protein:ligand down to 1:10.
>>
>>
>> Regards,
>>
>> George
>>
>> -----Original Message-----
>> From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of
>> Kavyashree Manjunath
>> Sent: Tuesday, February 09, 2016 7:30 AM
>> To: [log in to unmask]
>> Subject: [ccp4bb] Protein:ligand ratio for co-crystallization
>>
>> Dear users,
>>
>> I would like to know the minimum protein:ligand ratio that will yield
>> a cocrystal, provided there are some binding values from biophysical
>> techniques.
>>
>> Suppose, biophysical techniques like MST shows a binding when molar
>> ratio of protein:ligand is greater than 1:500, is it necessary to have
>> this ratio while co-crystallizing?
>>
>> I have previous experiences in which I had obtained ligand bound
>> structure when I used 1:10 to 1:30 (molar ratio). But, it is not working
> now.
>>
>> Thank you
>> Regards
>> Kavya
>>
>>
>> --
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>
>
>
>
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