Nitrilotriacetic acid-derivatized quantum dots for simple purification and site-selective fluorescent labeling of active proteins in a single step.
We demonstrate that QDs coated with nitrilotriacetic acid (NTA) bound to Ni (2+) can be used to reversibly and selectively bind, purify, and fluorescently label His 6-tagged (N-terminal) glutathione S-transferase (GST) in one step with retention of enzymatic activity. We find binding to be less effective in the absence of the His 6-tag or Ni (2+) ions.
Gupta
Manish
M
Caniard
Anne
A
Touceda-Varela
Angeles
A
Campopiano
Dominic J
DJ
Mareque-Rivas
Juan C
JC
D000818:Animals; D019187:Cadmium Compounds / Q000737:chemistry; D002264:Carboxylic Acids / Q000737:chemistry; D020134:Catalytic Domain; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D005982:Glutathione Transferase / Q000302:isolation & purification* / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D008958:Models, Molecular; D008968:Molecular Conformation; D009532:Nickel / Q000737:chemistry; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives*; D011485:Protein Binding; D045663:Quantum Dots; D012547:Schistosoma / Q000201:enzymology*; D018036:Selenium Compounds / Q000737:chemistry; D013379:Substrate Specificity; D013440:Sulfides / Q000737:chemistry; D013499:Surface Properties; D017967:Zinc Compounds / Q000737:chemistry
D019187:Cadmium Compounds; D002264:Carboxylic Acids; D005456:Fluorescent Dyes; D018036:Selenium Compounds; D013440:Sulfides; D017967:Zinc Compounds; D006639:Histidine; D009532:Nickel; C058667:cadmium selenide; D005982:Glutathione Transferase; D009571:Nitrilotriacetic Acid; C031238:zinc sulfide