single amino acid mutation effects on protein function - (Jan/03/2010 )

Hi
I have cloned a alpha subunit of translation initiation factor 2 for expression in mammalian cell, but in sequencing results there is a single amino acid substitution ( aspartic acid 301 in original sequence is substituted with glycin in my sequence) I dont know how harmfull is this substitution, how can I know this?
best

sara.r on Jan 3 2010, 07:14 PM said:

Hi Sara, ur mutation has led to replacement of an acidic residue with a neutral one....The effect of this substitution, according to me, will really depend on the role this amino acid has to play in ur protein's function.....in the case of enzymes, if this residue is involved in the catalytic site formation, u might see an effect.......its possible that it might not fold properly into its proper structure...........these are just speculations.....as far as I know, u really cannot predict the effect unless u purify and do an activity assay....of course, in many instances, u might not even see any effect.........Maybe u can check by multiple alignment if this is a conserved residue or something.....
all the best...