PMID- 10359649
OWN - NLM
STAT- MEDLINE
DCOM- 19990802
LR  - 20191210
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 340 ( Pt 3)
DP  - 1999 Jun 15
TI  - Protein modification during biological aging: selective tyrosine nitration of the
      SERCA2a isoform of the sarcoplasmic reticulum Ca2+-ATPase in skeletal muscle.
PG  - 657-69
AB  - The accumulation of covalently modified proteins is an important hallmark of
      biological aging, but relatively few studies have addressed the detailed
      molecular-chemical changes and processes responsible for the modification of
      specific protein targets. Recently, Narayanan et al. [Narayanan, Jones, Xu and Yu
      (1996) Am. J. Physiol. 271, C1032-C1040] reported that the effects of aging on
      skeletal-muscle function are muscle-specific, with a significant age-dependent
      change in ATP-supported Ca2+-uptake activity for slow-twitch but not for
      fast-twitch muscle. Here we have characterized in detail the age-dependent
      functional and chemical modifications of the rat skeletal-muscle
      sarcoplasmic-reticulum (SR) Ca2+-ATPase isoforms SERCA1 and SERCA2a from
      fast-twitch and slow-twitch muscle respectively. We find a significant
      age-dependent loss in the Ca2+-ATPase activity (26% relative to Ca2+-ATPase
      content) and Ca2+-uptake rate specifically in SR isolated from predominantly
      slow-twitch, but not from fast-twitch, muscles. Western immunoblotting and amino 
      acid analysis demonstrate that, selectively, the SERCA2a isoform progressively
      accumulates a significant amount of nitrotyrosine with age (approximately
      3.5+/-0. 7 mol/mol of SR Ca2+-ATPase). Both Ca2+-ATPase isoforms suffer an
      age-dependent loss of reduced cysteine which is, however, functionally
      insignificant. In vitro, the incubation of fast- and slow-twitch muscle SR with
      peroxynitrite (ONOO-) (but not NO/O2) results in the selective nitration only of 
      the SERCA2a, suggesting that ONOO- may be the source of the nitrating agent in
      vivo. A correlation of the SR Ca2+-ATPase activity and covalent protein
      modifications in vitro and in vivo suggests that tyrosine nitration may affect
      the Ca2+-ATPase activity. By means of partial and complete proteolytic digestion 
      of purified SERCA2a with trypsin or Staphylococcus aureus V8 protease, followed
      by Western-blot, amino acid and HPLC-electrospray-MS (ESI-MS) analysis, we
      localized a large part of the age-dependent tyrosine nitration to the sequence
      Tyr294-Tyr295 in the M4-M8 transmembrane domain of the SERCA2a, close to sites
      essential for Ca2+ translocation.
FAU - Viner, R I
AU  - Viner RI
AD  - Department of Pharmaceutical Chemistry, University of Kansas, Simons Building,
      2095 Constant Avenue, Lawrence, KS 66047, USA.
FAU - Ferrington, D A
AU  - Ferrington DA
FAU - Williams, T D
AU  - Williams TD
FAU - Bigelow, D J
AU  - Bigelow DJ
FAU - Schoneich, C
AU  - Schoneich C
LA  - eng
GR  - P01AG12993/AG/NIA NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Isoenzymes)
RN  - 0 (Nitrates)
RN  - 0 (Peptide Fragments)
RN  - 0 (Sulfhydryl Compounds)
RN  - 26404-66-0 (peroxynitric acid)
RN  - 31C4KY9ESH (Nitric Oxide)
RN  - 3604-79-3 (3-nitrotyrosine)
RN  - 42HK56048U (Tyrosine)
RN  - EC 3.4.- (Endopeptidases)
RN  - EC 7.2.2.10 (Calcium-Transporting ATPases)
RN  - K848JZ4886 (Cysteine)
RN  - S88TT14065 (Oxygen)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Aging/*metabolism
MH  - Amino Acid Sequence
MH  - Animals
MH  - Biological Transport
MH  - Calcium/metabolism
MH  - Calcium-Transporting ATPases/chemistry/*metabolism
MH  - Cysteine/metabolism
MH  - Endopeptidases/metabolism
MH  - Isoenzymes/chemistry/metabolism
MH  - Muscle Fibers, Fast-Twitch/enzymology
MH  - Muscle Fibers, Slow-Twitch/enzymology
MH  - Nitrates/metabolism
MH  - Nitric Oxide/metabolism
MH  - Oxygen/metabolism
MH  - Peptide Fragments/chemistry/metabolism
MH  - *Protein Processing, Post-Translational
MH  - Rats
MH  - Rats, Inbred F344
MH  - Sarcoplasmic Reticulum/*enzymology/metabolism
MH  - Sulfhydryl Compounds/metabolism
MH  - Tyrosine/*analogs & derivatives/analysis/*metabolism
PMC - PMC1220296
EDAT- 1999/06/09 00:00
MHDA- 1999/06/09 00:01
CRDT- 1999/06/09 00:00
PHST- 1999/06/09 00:00 [pubmed]
PHST- 1999/06/09 00:01 [medline]
PHST- 1999/06/09 00:00 [entrez]
PST - ppublish
SO  - Biochem J. 1999 Jun 15;340 ( Pt 3):657-69.