Site Specific Chemical Adduction to Cytochrome c
Chem. Res. Toxicol., Vol. 18, No. 1, 2005 49
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on the heterologous protein complex involving cytochrome
c and cytochrome b5 (31, 32). The key lysines involved in
the interaction are K13, K25, K27, K72, and K79. Thus,
preferential adduction of the K25-K27 region may result
in decreased binding and disruption of mitochondrial
respiration.
Release of cytochrome c and binding to apoptosis
protease activating factor-1 (Apaf-1) is an important
initiating event in the mitochondrial-controlled apoptotic
pathway (33, 34). Apaf-1 binding to cytochrome c has
been studied via a series of mutational epitopes, which
indicate that a number of residues are involved, including
K7, K25, K39, ETLM (62-65), and K72 (35). The forma-
tion of adducts at the K25 site could lead to disruption
of the Apaf-1:cytochrome c complex. Preliminary studies
have shown that adduction of cytochrome c by benzo-
quinone can inhibit initiation of apoptosis in cell lysates.2
Alkylating agents such as iodoacetamide produce less
specific protein adduction, which will less likely cause
cellular damage unless adducted sites critical for protein
function are present in high concentrations.
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modifications: New tools to study the effects of environmental
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Acknowledgment. Thanks go to Vanessa Recio for
purification of the GSHQ conjugate and Dr. Samy Habib
for conducting the initial reaction of GSBQ and cyto-
chrome c. Mass spectra for the cytochrome c experiments
were acquired in the Analytical Instrumentation Facility
Core of the Center for Research on Environmental
Disease (CRED) supported by NIEHS Grant ES07784.
This work was supported by grants from the National
Institutes of Health GM39338 and ES06694 to S.S.L.,
ES10056 and ES06694 to D.L., and a CRED Pilot Project
program grant to M.D.P. D.E.M. was supported by
NIEHS Training Grant ES07091.
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