Creatine Kinase and Creatine Transporter in Normal, Wounded, and Diseased Skin

Abstract

Skin comprises many cell types that are characterized by high biosynthetic activity and increased energy turnover. The creatine kinase system, consisting of creatine kinase isoenzymes and creatine transporter, is known to be important to support the high energy demands in such cells. We analyzed the presence and the localization of these proteins in murine and human skin under healthy and pathologic conditions, using immunoblotting and confocal immunohistochemistry with our recently developed specific antibodies. In murine skin, we found high amounts of brain-type cytosolic creatine kinase coexpressed with lower amounts of ubiquitous mitochondrial creatine kinase, both mainly localized in suprabasal layers of the epidermis, different cell types of hair follicles, sebaceous glands, and the subcutaneous panniculus carnosus muscle. With exception of sebaceous glands, these cells were also expressing creatine transporter. Muscle-type cytosolic creatine kinase and sarcomeric mitochondrial creatine kinase were restricted to panniculus carnosus. Immediately after wounding of murine skin, brain-type cytosolic creatine kinase and a creatine transporter-subspecies were transiently upregulated about 3-fold as seen in immunoblots, whereas the amount of ubiquitous mitochondrial creatine kinase increased during days 10-15 after wounding. Healthy and psoriatic human skin showed a similar coexpression pattern of brain-type cytosolic creatine kinase, ubiquitous mitochondrial creatine kinase, and creatine transporter in this pilot study, with creatine transporter species being upregulated in psoriasis.

Skin phosphocreatine

“…CRT is responsible for creatine uptake into the cell from the circulation. These Swiss investigators also observed increased CPK CRT activity in psoriasis samples (17), stating that their data seem to confirm previous reports of Zemtsov et al (17) on this subject. I could not agree more with the conclusion of this elegant study by Schlattner and Walliman: 'We detected the presence of functional PCR/CPK system in many metabolically very active cell types of skin, especially in epidermis and hair follicles.'…”

“…In 2002, Schlattner et al (17) observed in wounded murine skin a three-fold increase in CPK production: this makes sense as in injured skin, like in cardiac tissue after a myocardial infarction, there is an increase in CPK production, tissue damage and leakage of this enzyme into circulation. This is why every hospital, clinic, or laboratory is equipped to measure increased CPK levels after a myocardial infarction.…”

“…In 2002, an internationally renowned group of Swiss cell biologists (they wrote review articles on the subject of CPK enzymes) (8,9), utilizing molecular biology and immunohistochemical techniques, detected both CPK in the epidermis, hair follicles, and sebaceous glands (17). They also detected in these skin structures creatine transporter (CRT).…”

Acute toxicity of doxorubicin on isolated perfused heart: response of kinases regulating energy supply

“…Further dilutions were made in Krebs-Henseleit solution directly before heart perfusion. Primary antibodies against CK isoenzymes have been generated and characterized previously in our laboratory (39); other primary antibodies were commercial, including polyclonal anti-␣-AMPK (Cell Signaling), anti-phospho-(Thr 172 )-␣-AMPK (Cell Signaling), anti-acetyl-CoA carboxylase-␣ (ACC-␣; Alpha Diagnostics), anti-phospho(Ser 79 )-ACC-␣ (Upstate), anti-active caspase 3 (Cell Signaling), anti-cleaved poly(ADP-ribose) polymerase (PARP; BD Biosciences Pharmingen), anti-heart myosin binding protein C (gift of J. C. Perriard, Swiss Federal Institute of Technology), anti-voltage-dependent anion channel 1 (VDAC1; Santa Cruz Biotechnology), and anti-GAPDH (Santa Cruz Biotechnology) as well as monoclonal anti-cytochrome c (BD Biosciences Pharmingen), anti-sarcomeric myosin heavy chain (A41025; DSHB), and anti-␣ sarcomeric actinin (Ea-53, Sigma). The disodium salts of ADP and NADP were obtained from Roche, PCr from Calbiochem, and P 1 P 5 -di(adenosine-5Ј)-pentaphosphate pentasodium salt from Sigma.…”

Creatine kinase–mediated improvement of function in failing mouse hearts provides causal evidence the failing heart is energy starved

“…CK protein expression was determined by standard Western blotting techniques as previously reported (54)(55)(56). LV homogenates were separated on a 4%-12% Bis-Tris gradient gel under denaturing conditions as previously described (55,56) and using an anti-CK-M166-180 antibody produced in rabbit IgG fraction of antiserum and electroblotted onto nitrocellulose membrane (55,56).…”