biotinylated tag

Mikhail Alexeyev malexeye at JAGUAR1.USOUTHAL.EDU
Fri Nov 7 11:05:43 EST 1997


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 Joel Baguet wrote:

> Does anybody know how I can tag my protein with biotinylated amino
> acids in N terminal position.
>
> Is it a possibility with TNT promega kit with biotinylated leucin or
> methionin instead of radiolabelled leucin or methionin?
>
> or do I use tRNA for biotinylated leucin?
>
>
This does not precisely address your question, but biotinylation of
protein can be achieved by biotinmaleimide (modification of cysteine) or
by fusing your protein to biotin-acceptor domain from the oxaloacetate
decarboxylase of Klebsiella pneumoniae.  See for example:

Proc Natl Acad Sci U S A 1993 Aug 1;90(15):6934-6938

Properties and purification of an active biotinylated lactose
permease from Escherichia coli.

Consler TG, Persson BL, Jung H, Zen KH, Jung K, Prive GG, Verner GE,
Kaback HR

M.A.

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<HTML>
&nbsp;
<BR>&nbsp;Joel Baguet wrote:
<BLOCKQUOTE TYPE=CITE>
<PRE>
Does anybody know how I can tag my protein with biotinylated amino&nbsp;
acids in N terminal position.

Is it a possibility with TNT promega kit with biotinylated leucin or&nbsp;
methionin instead of radiolabelled leucin or methionin?

or do I use tRNA for biotinylated leucin?

</PRE>
</BLOCKQUOTE>
This does not precisely address your question, but biotinylation of protein
can be achieved by biotinmaleimide (modification of cysteine) or by fusing
your protein to biotin-acceptor domain from the oxaloacetate decarboxylase
of Klebsiella pneumoniae.&nbsp; See for example:

<P>Proc Natl Acad Sci U S A 1993 Aug 1;90(15):6934-6938

<P>Properties and purification of an active biotinylated lactose
<BR>permease from Escherichia coli.

<P>Consler TG, Persson BL, Jung H, Zen KH, Jung K, Prive GG, Verner GE,
Kaback HR
<BR>&nbsp;
<BR>M.A.</HTML>

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