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Surface-assisted de...
Surface-assisted delivery of fluorescent groups to hGST A1-1 and a lysine mutant
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- Viljanen, Johan (författare)
- Linköpings universitet,Organisk Kemi,Tekniska högskolan
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- Tegler, Lotta (författare)
- Linköpings universitet,Organisk Kemi,Tekniska högskolan
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- Larsson, J. (författare)
- Linköpings universitet,Organisk Kemi,Tekniska högskolan
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- Broo, Kerstin, 1970 (författare)
- Linköpings universitet,Gothenburg University,Göteborgs universitet,Institutionen för kemi,Department of Chemistry,Organisk Kemi,Tekniska högskolan
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(creator_code:org_t)
- American Chemical Society (ACS), 2006
- 2006
- Engelska.
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Ingår i: Bioconjugate Chemistry. - : American Chemical Society (ACS). - 1043-1802 .- 1520-4812. ; 17:2, s. 429-437
- Relaterad länk:
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http://urn.kb.se/res...
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https://gup.ub.gu.se...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Human glutathione transferase (hGST) Al-l and a lysine mutant(A216K) can both be rapidly and site-specifically acylated on Y9 and K216, respectively, using a range of thiolesters of glutathione (GS-thiolesters) as modifying reagents. The present investigation was aimed at developing a method with which to deliver a fluorescent acyl group from a solid support under conditions compatible with standard protein purification schemes. A number of fluorescent GS-thiolesters with modified peptide backbones were therefore prepared and tested for reactivity toward hGST Al-l and the A216K mutant. Substitutions at the alpha-NH2 part of the glutathione backbone were not tolerated by the proteins. However, two fluorescent reagents that carry a biotin moiety at the C-terminal part of glutathione were found through MALDI-MS experiments to react in solution with Y9 of the wild-type protein and one reagent with K216 of A216K. The reaction can take place in the presence of glutathione and even in a crude E. coli lysate of cells expressing A216K. Delivery of the fluorescent group to Y9 or K216 was possible using, NeutrAvidin (NA) beads that had been preincubated with biotinylated reagent. Alternatively, excess reagent can be removed by a brief incubation with NA beads. We have thus now developed a system for protein labeling with easy removal of excess and used up low-molecular weight reagent. This strategy can conceivably be utilized in future protein purification and labeling experiments.
Ämnesord
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
Nyckelord
- GLUTATHIONE-S-TRANSFERASE
- PROTEIN EXPRESSION
- HUMAN GENOME
- ISOENZYMES
- EVOLUTION
- ANALOGS
- PEPTIDES
- ENZYMES
- RESIDUE
- GLYCYL
- human GST A1-1
- NATURAL SCIENCES
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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