New markers of oxidative damage to macromolecules
JMB 2008; 27 (1)
DOI: 10.2478/v10011-007-0049-x
UDK 577.1 : 61
ISSN 1452-8258
JMB 27: 1–16, 2008
Review article
Pregledni ~lanak
NEW MARKERS OF OXIDATIVE DAMAGE TO MACROMOLECULES
NOVI MARKERI OKSIDATIVNOG O[TE]ENJA MAKROMOLEKULA
Emina ^olak
Institute of Medical Biochemistry, CCS, Belgrade, Serbia
Summary: The presence of free radicals in biological material has been discovered some 50 years ago. In physiological
conditions, free radicals, in the first place the ones of oxygen
and nitrogen, are continuously synthesized and involved in
the regulation of a series of physiological processes. The
excess of free radicals is efficiently eliminated from the body
in order to prevent their toxic effects. Toxic effects of free
radicals may be classified into three groups: a) change of
intracellular redox potential, b) oxidative modification of lipids, proteins and DNA, and c) gene activation. Lipid peroxidation involving cell membranes, lipoproteins and other
molecules leads to the production of primary high-reactive
intermediaries (alkyl radicals, conjugated dienes, peroxy- and
alkoxyl radicals and lipid hydroperoxide), whose further
breakdown generates the secondary products of lipid peroxidation: short-chain evaporable hydrocarbons, aldehydes and
final products of lipid peroxidation: isoprostanes, MDA, 4hydroxy-2,3-transnonenal and 4,5-dihydroxydecenal which
are important mediators of atherosclerosis, coronary disease,
acute myocardial infarction, rheumatoid arthritis, systemic
sclerosis and lupus erythematodes. Oxidative modification of
proteins is manifested by changes in their primary, secondary
and tertiary structures. Proteins have a specific biological
function, and therefore their modification results in unique
functional consequences. The nature of protein modification
may provide valid information on the type of oxidants causing
the damage. Chlorotyrosyl is a specific marker of oxidative
damage to tyrosine caused by HOCl action, which most
commonly reflects the involvement of neutrophils and monocytes in oxidative stress, while nitrotyrosyl indicates the presence of higher peroxy-nitrite synthesis. Methyonin and cysteine are the amino acids most sensitive to oxidative stress,
carbonyl groups are markers of severe damage caused by
free radicals, and di-tyrosyl is the most significant and sensitive marker of oxidative modification made by g rays. »Carbo-
Address for correspondence:
^olak Emina
Institute of Medical Biochemistry
Clinical Center of Serbia
Vi{egradska 26
11000 Belgrade, Serbia
tel: 3615 631
e-mail: eminacolakªsbb.co.yu
Kratak sadr`aj: Prisustvo slobodnih radikala u biolo{kim
materijalima je otkriveno pre ne{to vi{e od 50 godina. U
fiziolo{kim uslovima, slobodni radikali, pre svega kiseonika
i azota, stalno se sinteti{u i pri tom u~estvuju u regulaciji
niza fiziolo{kih procesa. Vi{ak slobodnih radikala se efikasno
uklanja iz organizma u cilju spre~avanja njihovih toksi~nih
efekata. Toksi~ni efekti delovanja slobodnih radikala se mogu podeliti u tri grupe: a) pomeranje intra}elijskog redoks
stanja, b) oksidativna modifikacija lipida, proteina i DNK i
c) aktivacija gena. Lipidna peroksidacija koja zahvata }elijske membrane, lipoproteine i druge molekule dovodi do
stvaranja primarnih visokoreaktivnih intermedijera (alkil
radikala, konjugovanih diena, peroksi i alkoksi radikala i lipidnih hidroperoksida), koji daljom razgradnjom daju sekundarne produkte lipidne peroksidacije: kratkolan~ane
isparljive ugljovodonike, aldehide i krajnje proizvode lipidne
peroksidacije: izoprostane, MDA (malondialdehid), 4-hidroksi-2,3-transnonenal i 4,5-dihidroksidecenal, koji su
zna~ajni medijatori ateroskleroze, koronarne bolesti, akutnog infarkta miokarda, reumatoidnog artritisa, sistemske
skleroze, lupusa eritematodesa. Oksidativna modifikacija
proteina manifestuje se promenama njihove primarne, sekundarne i tercijarne strukture. Proteini imaju specifi~nu
biolo{ku funkciju, stoga njihova izmena ima jedinstvene
funkcionalne posledice. Priroda proteinske modifikacije
mo`e nam dati validan podatak o vrsti oksidansa koji je
doveo do o{te}enja. Hlorotirozil je specifi~an marker oksidativnog o{te}enja tirozina dejstvom hipohlorita (HOCl),
koji naj~e{}e reflektuje u~e{}e neutrofila i monocita u oksidativnom stresu, dok nitrotirozil ukazuje na prisustvo pove}ane sinteze peroksinitrita. Metionin i cistein su najosetljivije aminokiseline na dejstvo oksidativnog stresa, karbonilne
grupe su markeri te{kog o{te}enja slobodnim radikalima,
dok je di-tirozil najzna~ajniji i najosetljiviji marker oksidativne modifikacije dejstvom g-zraka. »Karbonilni stres« je zna~ajan vid sekundarne oksidacije proteina u kojem redukuju}i {e}eri neenzimski reaguju sa amino grupama proteina
i lipida dovode}i do stvaranja kovalentnih jedinjenja poznatih kao uznapredovani proizvodi glikozilacije ili AGE-proizvodi (advanced glycosylated end products). Hidroksilni radikal o{te}uje i molekul DNK dovode}i do gubitka baze i
stvaranja abazi~nih mesta (AP site-ova), cepanja lanca
DNK i modifikacije {e}era. Krajnji proizvodi lipidne peroksidacije (MDA) mogu se kovalentno vezivati za DNK, stvara-
2 ^olak: New markers of oxidative damage to macromolecules
nyl stress« is an important form of the secondary oxidation of
proteins, where reducing sugars non-enzymatically react with
amino groups of proteins and lipids and give rise to the production of covalent compounds known as advanced glycosylated end products (AGE-products). A hydroxyl radical
damages the DNA, leading to a loss of base and the formation of abasic sites (AP sites), break of DNA chain and sugar
modification. Final lipid peroxidation products (MDA) may
covalently bind to DNA, producing the »DNA radicals« which
are responsible for mutations. Measurement of an adequate
oxidative stress biomarker may not only point to an early
onset of disease, its progression and assessment of therapy
effectiveness, but can also help in the clarification of the
pathophysiological mechanisms of tissue damage caused by
oxidative stress, prediction of disease prognosis and choice of
appropriate treatment in the early stages of disease.
ju}i »DNK radikale« koji su odgovorni za nastanak mutacija. Merenje adekvatnog biomarkera oksidativnog stresa
mo`e nam ukazati ne samo na ranu pojavu bolesti, njenu
progresiju i procenu efikasnosti terapije, ve} i pomo}i u
rasvetljavanju patofiziolo{kih mehanizama o{te}enja tkiva
dejstvom oksidativnog stresa, u predikciji prognoze bolesti
i izboru adekvatnog le~enja u ranim stadijumima bolesti.
Klju~ne re~i: oksidativna modifikacija, lipidi, proteini, DNK,
biomarkeri, slobodni radikali, lipidna peroksidacija, karbonilni
stres
Keywords: oxidative modification of lipids, proteins, DNA,
biomarkers, free radicals, lipid peroxidation, carbonyl stress
Introduction
The presence of free radicals in biological materials was discovered some 50 years ago (1). Soon
after that discovery, Harman (2) reported his hypothesis that free radicals were the (...truncated)