Hi Harshitha,
What you could do is to insert a big loop between your protein and the tag.
Once you have other crystal structures to compare where it interacts, this
should give you clue if it is also better to tag the N or C terminus.
Best wishes
______________________________________________________
Rafael Marques da Silva
PhD Student – Structural Biology
University of Leicester
Mestre em Física Biomolecular
Universidade de São Paulo
Bacharel em Ciências Biológicas
Universidade Federal de São Carlos
phone: +44 07861 273773
"A sorte acompanha uma mente bem treinada"
________________________________________________
________________________________
De: CCP4 bulletin board <[email protected]> em nome de Harshitha H N
<[email protected]>
Enviado: segunda-feira, 6 de julho de 2026 05:53
Para: [email protected] <[email protected]>
Assunto: [ccp4bb] Need advice on designing bacterial protein co-localization
experiments.
I am designing a bacterial protein interaction and co-localisation study
involving two interacting proteins.
One protein is very small (~7–8 kDa), while the other is approximately 25–30
kDa. Previous biochemical experiments (crystal structure and binding studies)
have confirmed that the two proteins interact, and one protein modulates the
function of the other.
My initial plan was to generate fluorescent protein fusions (EGFP and mCherry)
for live-cell co-localisation. However, I am concerned that fusing a ~27–29 kDa
fluorescent protein to the smaller protein may interfere with its folding,
localisation, or interaction.
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