To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Heme incorporation in expressed protein
Dear all,
Sorry for non-crystallography related question.
We are trying to express an eukaryotic heme protein in E. coli. We are able to
express it in soluble form. When we use 5-Aminolevulinic Acid, E. coli becomes
I was able to express a heme protein by inducing and expressing at
room temperature and using a promoter weaker than T7 (can't remember
the exact one right now). The key was to slow down the rate of protein
production to allow heme incorporation. You might try using less IPTG
too.
Ho Leung Ng
Incomplete heme-incorporation in heme proteins expressed in E. coli. is a
common issue. Check out the following paper for an easy method, which, in
our experience, has solved all our issues regarding this problem. This has
worked really well for many different heme-binding proteins (Cys / His
Dear all,
Sorry for non-crystallography related question.
We are trying to express an eukaryotic heme protein in E. coli. We are able to
express it in soluble form. When we use 5-Aminolevulinic Acid, E. coli becomes
brown. However, after cell lysis the soluble protein contains no heme and the
