BIOC2000 Study Guide - Final Guide: Green Fluorescent Protein, Molecular Mass, Glutathione
GFP
>sp|P42212|GFP_AEQVI Green fluorescent protein OS=Aequorea victoria OX=6100 GN=GFP PE=1 SV=1
MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTL
VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLV
NRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLAD
HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
GST
>sp|P08515|GST26_SCHJA Glutathione S-transferase class-mu 26 kDa isozyme OS=Schistosoma japonicum OX=6182 PE=1
SV=3
MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYID
GDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKV
DFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFK
KRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
GFP mutant
>sp|P42212|GFP_AEQVI Green fluorescent protein OS=Aequorea victoria OX=6100 GN=GFP PE=1 SV=1
MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTL
VTTFGYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLV
NRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLAD
HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
GST-GFPS65T Fusion
MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYID
GDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKV
DFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFK
KRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKMSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGVQCFSRYPD
HMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGD
GPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
Molecular Weight(MW) and PI of GST, GFPS65T, GST-GFPS65T fusion
GFP:
Number of amino acids: 238
Molecular weight: 26886.32
Theoretical pI: 5.67
GST:
Document Summary
>sp|p08515|gst26_schja glutathione s-transferase class-mu 26 kda isozyme os=schistosoma japonicum ox=6182 pe=1. >sp|p42212|gfp_aeqvi green fluorescent protein os=aequorea victoria ox=6100 gn=gfp pe=1 sv=1. Molecular weight(mw) and pi of gst, gfps65t, gst-gfps65t fusion. Protein denature because they lose their shape from heat increasing kinetic energy which breaks weak intermolecular bonds like hydrogen bonds and protein secondary structure. Protein however will not be degraded since backbone is held together by strong covalent bond which is 10 times harder to break. Hydrogen bond drives dna base pairing and it determines the shape of the molecule. At phpka, the deprotonated form will be more common (a- or b) At ph=pka, there will be equal concentrations of each form. N terminus pka =9. 6 and c terminus pka=2. When ph is at 7 in the case of n terminus, ph is lower than pka so more nh3+ form,
