Anti‐Oxidative Response of Carbonic Anhydrase III in Skeletal Muscle

Abstract

SummaryWe propose that carbonic anhydrase III (CAIII) functions as an anti-oxidant agent in skeletal muscle. To explore this hypothesis, we analyzed the gene expression profile of skeletal muscle in mice deficient in CAIII gene utilizing the murine genome U74Av2 set microarray. Pairwise comparison between CAIII knockout mice and their wild-type littermates revealed that more than 500 of 12,000 genes in the array showed an altered level of transcription. Of particular note were transcriptional alterations among genes associated with the glutathione redox cycle, suggesting a possible involvement of CAIII in the glutathione-mediated anti-oxidant activity. We therefore investigated S-glutathiolation and irreversible oxidation of the 2 reactive sulfhyryls of CAIII in skeletal muscle under oxidative stresses of ischemia, or exhaustive exercise. Analysis by isoelectric focusing followed by Western blot revealed that the two sulfhydryls were differentially and progressively oxidized. Brief ischemia of 10 -20 min provoked partial (one of the suflhydryls) modification of CAIII via reversible S-glutathiolation. Protracted 60 min ischemia yielded equal amounts of both partially and completely (both sulfhydryls) modified CAIII due to irreversible oxidization. Twenty minutes of repetitive electrical stimulation, simulating exhaustive exercise, produced a mixed yield: partial modification by reversible Sglutathiolation and complete modification by irreversible oxidation. Thus CAIII responds to oxidative stress with a distinctive sulfhydryl oxidation patterns reflecting duration and severity that may prove sensitive indices of extent and type of damage in muscle injury.

Carbonic anhydrase III (Car3) is not required for fatty acid synthesis and does not protect against high-fat diet induced obesity in mice

“…S-glutathionylation of proteins is an early response to oxidative damage [77]. S-glutathionylation of Car3 increases with increased oxidative damage that occurs naturally as in aging [78] or with muscle damage [22]. Car3 contains numerous other additional cysteine residues not found in Car1/2.…”

“…However, CA III does differ in amino acid alignment with the closely related CA I and CA II proteins at two important residues, as depicted in (Fig 1), lysine 64 (Lys) and phenylalanine 198 (Phe), which dramatically decrease the activity of CA III [13–17]. While the function of CA III is currently unknown it has been speculated to be involved in metabolism, oxidative damage response, and mitochondrial ATP synthesis [18–22]. …”

“…No obvious deleterious phenotypes have been reported in previous studies on Car3 knockout mice; however, the effect of specialized diets on these knockout mice was not previously researched [19, 20, 22, 49, 50]. In this study, we investigate Car3 function in de novo lipogenesis by measuring fatty acid synthesis in isolated mouse hepatocytes, and during induced obesity through high-fat diet by measuring subsequent changes in body weight, dyslipidemia, insulin resistance, food intake, and energy expenditure.…”

Multivariate Modeling of Proteins Related to Trapezius Myalgia, a Comparative Study of Female Cleaners with or without Pain

“…Carbonic anhydrase III is expressed predominantly in skeletal muscle and has been suggested to be an indicator of muscle damage [39]. It has been proposed that this protein functions as an essential anti-oxidant agent in skeletal muscle [40]. Decreased levels of carbonic anhydrase III in the myalgic muscle may indicate an imbalance in the cellular redox potential in this condition.…”

Ectopic expression of mitochondrial gamma carbonic anhydrase 2 causes male sterility by anther indehiscence

“…CA III is a cytosolic enzyme highly expressed in skeletal muscle in rats. It was demonstrated that in skeletal muscle, CA III also exhibits a protective role against peroxide (Zimmerman et al 2004). Thus, the complex I integrated CAs could protect tissues against peroxides when its normal expression is high.…”