interference between his-tags and signal factors

Kyle Legate legatek at mcmail.cis.mcmaster.ca
Tue Jul 16 11:11:19 EST 2002


On 16 Jul 2002, Emma Master wrote:

> I am trying to express a plant protein in Pichia. I have included a his-tag
> at the N-terminus of the expressed protein, which will occur between the
> protein of interest and a signal factor for secretion of the protein to the
> supernatant. Is it probable that the his-tag will interfere with cellular
> recognition of the signal factor, and thereby inhibit protein secretion?
>
There will be no problem. Signal sequences are stretches of hydrophobic
residues that while present at the amino terminus, can be flanked on each
side by non-hydrophobic residues. Bear in mind that many signal sequences
from secreted proteins are cleaved by signal peptidase as they cross the
ER membrane. You must ensure the signal peptidase cleavage site is absent
from your construct or your His tag will be removed!

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