Problems in N-terminal tagging

Tarja Kokkola tkokkola at
Mon Nov 26 14:57:25 EST 2001

Dear fellow bio-scientists,

Is it necessary to add any signal peptide sequences or a Kozak
sequence when
building a N-terminally tagged G protein-coupled receptor construct?
By now,
I have always just added a tag sequence after the initiation codon of
receptor and have had success in expressing the tagged receptor in
cells. There has been no consensus Kozak nucleotide sequence before
initiation codon, either.

I heard that membrane proteins need a N-terminal signal peptide which
them to the plasma membrane and that a N-terminal tag would be cut off
if it had been inserted before that peptide (because the signal
peptide is cleaved off during the transport process). Can this be true
for some GPCRs??

Please help me clarify that problem. Some Internet links regarding
N-terminal signal sequences would be welcome, too.

Tarja Kokkola

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