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Volumn 2, Issue 3, 2011, Pages 511-520

Mechanisms of chemical protein 19F-labeling and NMR-based biosensor construction in vitro and in cells using self-assembling ligand-directed tosylate compounds

Author keywords

[No Author keywords available]

Indexed keywords

AFFINITY LIGANDS; BIOCHEMICAL STUDIES; BIOLOGICAL RESEARCH; CHEMICAL LABELING; DRUG DISCOVERY; IN-CELL; IN-VITRO; LABELING REAGENT; LIGAND BINDING; NEW OPPORTUNITIES; NONCOVALENT PROTEINS; NOVEL PROTEINS; POTENTIAL UTILITY; PROTEIN LABELING; RATIONAL DESIGN; RED BLOOD CELL; SELF-ASSEMBLED; SELF-ASSEMBLING; SYNTHETIC PROBES; TARGET PROTEINS;

EID: 79955675069     PISSN: 20416520     EISSN: 20416539     Source Type: Journal    
DOI: 10.1039/c0sc00513d     Document Type: Article
Times cited : (37)

References (67)
  • 33
    • 70149112322 scopus 로고    scopus 로고
    • A recent example in which the basic strategy is likely to be applicable to cellular systems
    • A recent example in which the basic strategy is likely to be applicable to cellular systems: M. A. Brun, K.-T. Tan, E. Nakata, M. J. Hinner and K. Johnsson, J. Am. Chem. Soc., 2009, 131, 5873-5884.
    • (2009) J. Am. Chem. Soc , vol.131 , pp. 5873-5884
    • Brun, M.A.1    Tan, K.-T.2    Nakata, E.3    Hinner, M.J.4    Johnsson, K.5
  • 52
    • 79955668377 scopus 로고    scopus 로고
    • The 19F NMR signal of 1 was silent with CAI and EZA over 24 h in vitro
    • The 19F NMR signal of 1 was silent with CAI and EZA over 24 h in vitro.
  • 53
    • 79955670299 scopus 로고    scopus 로고
    • Chemically synthesized FBOH in buffer solution also showed a signal at -62.9 ppm (Fig. S2, ESI†)
    • Chemically synthesized FBOH in buffer solution also showed a signal at -62.9 ppm (Fig. S2, ESI†).
  • 54
    • 79955669757 scopus 로고    scopus 로고
    • Note
    • Since we set the experimental conditions where all of the labeling reagents are complexed with CAI, the determined values of kL and kHcomplex are considered as the rates from the complexed state between CAI and the reagents.
  • 55
    • 79955672350 scopus 로고    scopus 로고
    • Note
    • It should be noted that because of the need for a relatively high concentration of FB-CAI (20 mM) for 19F NMR measurements, we were unable to determine the precise affinity values of FB-CAI towards the strong inhibitors, EZA, AAZ and NBS.
  • 56
    • 79955649949 scopus 로고    scopus 로고
    • Note
    • The labeling site of FB-eCAI was shown to be at His67, which is identical to the site identified for FB-CAI prepared in vitro.
  • 57
    • 79955647589 scopus 로고    scopus 로고
    • Note
    • Under the conditions using less than 400 M of 1, we did not observe any toxicity for RBCs over the time period of the assay (over 48 h).
  • 58
    • 79955655042 scopus 로고    scopus 로고
    • Note
    • The validity of this mole number was also supported by in-cell 19F NMR measurements, in which the intracellular concentration of 1 can be determined from an integral of the signal at -62.3 ppm corresponding to the 1-eCAI complex (Fig. 4b). The mole number estimated in this manner was 194 ± 2 nmol.
  • 61
    • 79955647411 scopus 로고    scopus 로고
    • Note that following mixing of the RBC suspension with 1, the supernatant is replaced with fresh buffer before starting incubation (also see the Methods section)
    • Note that following mixing of the RBC suspension with 1, the supernatant is replaced with fresh buffer before starting incubation (also see the Methods section).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.