Re: [ccp4bb] co-crystallization

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

Xtal community:

- CCP4BB

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: co-crystallization
From: yangliuqing yangliuqing1984 {- at -} HOTMAIL {- dot -} COM
Date: 2008-12-01

Next message:
Subject: Re: co-crystallization
From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU
Date: 2008-12-01

Subject: Re: co-crystallization
From: mesters mesters {- at -} BIOCHEM {- dot -} UNI-LUEBECK {- dot -} DE
Date: 2008-12-01

Yes!, there is:

the fraction of occupied protein with substance can be calculated: S /
(S + Km) with S being the concentration of the compound.

So, if S = Km, half of the sites are occupied (it follows from
Michaelis-Menten theory).

In order to saturate the enzyme for 90,90909 % with the compound:

1) S = 10 x Kd (concentration of S at least 10 times the Kd)
and
2) S > P (total concentration of S must be larger than total
concentration of protein or "binding sites")

Depending on the solubility of the compound, this is not always
possible. In such a case, you need to use DMSO and/or add solid compound
to the protein solution and leave it for quite some time for the
compound to finally bind to the protein.

- J. -

yangliuqing wrote:
> Hello,everyone,
> I have a question for cocrystallization, is there some relationship
> between Km value and substrate concentration when making
> cocrystallization? How can I know the substrate is enough for binding?
> Thank you very much!
> liuqing
>
> ------------------------------------------------------------------------
> å…«å¦å¨±ä¹åŒ…æ‰“å¬ï¼ŒMSNèµ„è®¯é€Ÿé€’å¸®ä½ å¿™ï¼ äº†è§£è¯¦ç»†ï¼
>

--
Dr. Jeroen R. Mesters
Gruppenleiter Strukturelle Neurobiologie und Kristallogenese
Institut fÃ¼r Biochemie, UniversitÃ¤t zu LÃ¼beck
Zentrum fÃ¼r Medizinische Struktur- und Zellbiologie
Ratzeburger Allee 160, D-23538 LÃ¼beck
Tel: +49-451-5004065, Fax: +49-451-5004068
Http://www.biochem.uni-luebeck.de
Http://www.iobcr.org
Http://www.selfish-brain.org
Http://www.opticryst.org
--
If you can look into the seeds of time and say
which grain will grow and which will not - speak then to me (Macbeth)
--

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: co-crystallization
From: yangliuqing yangliuqing1984 {- at -} HOTMAIL {- dot -} COM
Date: 2008-12-01

Next message:
Subject: Re: co-crystallization
From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU
Date: 2008-12-01