OtherBasic Science Investigations

131I-Labeled Peptides as Caspase Substrates for Apoptosis Imaging

Claudia Bauer, Ulrike Bauder-Wuest, Walter Mier, Uwe Haberkorn and Michael Eisenhut
Journal of Nuclear Medicine June 2005, 46 (6) 1066-1074;

Article Figures & Data

Figures

  • FIGURE 1.
    FIGURE 1.

    Time-course development of apoptosis obtained with camptothecin-treated Jurkat cells. (A and B) Flow cytometry using Alexa Fluor 488-labeled annexin V showing phosphatidylserine externalization (A) and formation of fluorescent rhodamine 110 mediated through caspase-3 hydrolysis of Z-DEVD-R110 (B) (n = 3). Data are expressed as mean ± SD. arb. units = arbitrary units.

  • FIGURE 2.
    FIGURE 2.

    Propidium iodide (FL-III)/Alexa Fluor 488-annexin V (FL-I) flow cytometry of Jurkat cells treated with 10 μmol/L camptothecin for 4 h (A) and 1 μmol/L staurosporine for 3 h (B). Q1 = apoptotic cells; Q2 = apoptotic and necrotic cells; Q3 = living cells; Q4 = dead cells.

  • FIGURE 3.
    FIGURE 3.

    Uptake kinetics of 99mTc-HYNIC-annexin V in apoptotic and control cells. Cells showed 62% apoptosis with Alexa Fluor 488-annexin V measured by flow cytometry (n = 3). Data are expressed as mean ± SD.

  • FIGURE 4.
    FIGURE 4.

    Uptake in apoptotic and control cells of 131I-labeled DEVDGY-NH2 (1) and YDEVDG-NH2 (2) (A) as well as NQVNGY-NH2 (3) and YNQVNG-NH2 (4) (B) (n = 3). Data are expressed as mean ± SD.

  • FIGURE 5.
    FIGURE 5.

    Uptake in apoptotic and control cells of yDEVDG-conjugated Tat sequences (5–10) (n = 3). Data are expressed as mean ± SD.

  • FIGURE 6.
    FIGURE 6.

    Rhodamine 110 formation through caspase-mediated hydrolysis of Z-DEVD-R110 with increasing amounts of peptides 2 and 7 (n = 3). Data are expressed as mean ± SD.

  • FIGURE 7.
    FIGURE 7.

    MALDI mass spectra of peptide 7 incubated with (A) and without (B) commercial caspase-3 (m/z represents molecular mass/charge).

Tables

  • TABLE 1

    Peptides Used for Uptake Studies in Apoptotic and Control Cells

    PeptideSequenceMolecular weightMALDI-MS
    1DEVDGY-NH2695.7718.6 [M+Na]+
    2YDEVDG-NH2695.7696.8 [M+H]+
    3NQVNGY-NH2692.7715.8 [M+Na]+
    4YNQVNG-NH2692.7715.7 [M+Na]+
    5yDEVDG-Tat49–57-NH22,017.42,017.7 [M+H]+
    6yDEVDG-Tat57–49-NH22,017.42,018.4 [M+H]+
    7Tat49–57-yDEVDG-NH22,017.42,017.7 [M+H]+
    8Tat57–49-yDEVDG-NH22,017.42,016.9 [M+H]+
    9tat49–57-yDEVDG-NH22,017.42,018.4 [M+H]+
    10tat57–49-yDEVDG-NH22,017.42,017.7 [M+H]+
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