Toxicant‐induced alterations in two‐dimensional electrophoretic patterns of hepatic and renal stress proteins

Abstract

Recent studies in this laboratory and by others suggest that two-dimensional polyacrylamide gel electrophoresis of proteins (2-DE) possesses significant utility in the detection of chemical toxicity and in providing information regarding toxic mechanism. After having identified a set of specific heat-shock and glucose-regulated proteins whose expression in rodent liver and kidney is highly conserved and constitutive, we compared the effect of in vivo exposure to perfluoro-n-octanoic acid and perfluoro-n-decanoic acid on their expression. The following stress proteins were identified, their x, y coordinate positions mapped, and abundance statistically analyzed and compared: hsp32, hsp60, hsc70, hsp70, hsp90, grp75, grp94, protein disulfide isomerase (PDI), and ER60. We report here that the stress response to perfluorocarboxylic acids is tissue-, toxicant-, and stress protein class-specific and dose-related. Furthermore, because nearly all of the proteins studied were constitutively expressed at detectable levels in both liver and kidney, the 2-DE stress protein pattern may be suitable to future toxicologic screening applications.

Identification of Oxidative Stress-Related Proteins for Predictive Screening of Hepatotoxicity Using a Proteomic Approach

“…Toxicant-induced alterations of stress-related proteins have been previously reported (Witzmann et al, , 1996. In the present study, several oxidative stress-related proteins were affected by more than one hepatotoxicant.…”

Heat shock proteins in human cancer

“…The hsp70 chaperones increase resistance to cell death induced by TNF, monocytes, oxidative stress, ceramide, terminal differentiation, UV radiation, caspase-3 overexpression, ischemic injury, sepsis-induced injury, hydrogen peroxide toxicity, apoptosis, nitric oxide acid agents, and toxic drugs [63,64]. Inhibition of hsp70 expression diminishes cell survival, resulting in human fibroblasts treated with anti-hsp70 antibodies and subjected to sublethal temperatures [65].…”

Protein Markers of Chemical Exposure and Molecular Epidemiology

“…Because the cortex is the primary site of filtration and filtrate reabsorption and the medulla is the site of renal filtrate concentrating mechanisms, it's not surprising to find that the two regions are constitutively different [12,13] and that these regions are differentially susceptible to toxic exposure [14,15]. Examining the effect of xenobiotic exposure on the expression of whole kidney homogenate protein patterns, as we have done previously [16,17], may not accurately reflect potential regiospecific changes. This is particularly true if these protein alterations are regionally diametrical, and thus may actually minimize or mask one another and be overlooked in analyzing a whole tissue homogenate.…”

“…Animals were dosed on a mg/kg basis as follows: 260, 360, or 1300 ug/kg lead to achieve targeted blood levels of 20, 40 or 80 ug/dL. Following the loading dose period, rabbits were then placed on a maintenance dose solutions during study weeks [11][12][13][14][15][16][17][18][19][20] Dayton OH. Warm and cold ischemic times were 10 and 50 min, respectively.…”