Mechanisms of Ageing and Development
107 (1999) 245–253
Carbonylated proteins in aging and exercise:
immunoblot approaches
S. Goto
a,
*, A. Nakamura
a
, Z. Radak
a,b
, H. Nakamoto
a
,
R. Takahashi
a
, K. Yasuda
c
, Y. Sakurai
a,c
, N. Ishii
c
a
Department of Biochemistry, School of Pharmaceutical Sciences, Toho Uniersity, Miyama, Funabashi,
Chiba 274 -8510, Japan
b
Hungarian Uniersity of Physical Education, Alkotas u. 44, H-1123 Budapest, Hungary
c
Department of Molecular Life Science, Tokai Uniersity School of Medicine, Isehara 257 -0811, Japan
Received 1 May 1998; received in revised form 6 August 1998; accepted 26 September 1998
Abstract
Protein carbonyls were studied in aging and exercise by immunoblot followed by one- or
two-dimensional polyacrylamide gel electrophoresis using antibodies against 2,4-dinitro-
phenylhydrazones. Proteins of rat kidneys exhibited significant age-related increase in the
amount of carbonyl while those of the brain and liver did not. Major carbonylated proteins
in the kidney included serum albumin. In nematodes in which protein carbonyls increased
with age, one of the carbonylated proteins was identified as vitellogenin, an egg-yolk protein.
A possible biological significance of this protein present in abundance even after egg-laying
stages is discussed in terms of protection against oxidative stress. Exhaustive exercise induced
significant increase in the carbonylation of selected but unidentified proteins in the lung. This
oxidative stress might be caused by xanthine oxidase in this tissue and hypoxanthine derived
from ATP-depleted muscles. Exercise at high altitude caused higher carbonylation of the
skeletal muscle proteins, most notably a protein likely to be actin, than that at sea level but
no significant difference was observed in lipid peroxidation. These studies emphasize the
value of immunoblot analysis of tissue protein carbonyls in a variety of situations where
oxidative stress is likely involved. © 1999 Elsevier Science Ireland Ltd. All rights reserved.
* Corresponding author. Tel.: +81-474-721531; fax: +81-474-721531.
E-mail address: goto@phar.toho-u.ac.jp (S. Goto)
0047-6374/99/$ - see front matter © 1999 Elsevier Science Ireland Ltd. All rights reserved.
PII:S0047-6374(98)00133-X