Epidermal purification

Yada, Y., Higuchi, K., and Imokawa, G., 1995, Purification and biochemical characterization of membrane-bound epidermal ceramidases from guinea pig skin. J. Biot. Chem. 270 12677-12684. 

Bernard, D., Mehul, B., Delattre, C., Simonetti, L., Thomas-Collignon, A., and Schmidt, R. Purification and characterization of the endoglycosidase heparanase 1 from human plantar stratum corneum a key enzyme in epidermal physiology, J. Invest. Dermatol., 117, 1266-1273, 2001. 

Cohen, S., Carpenter, G., and King, L., Jr. (1980). Epidermal growth factor-receptor-protein kinase interactions. Co-purification of receptor and epidermal growth factor-enhanced phosphorylation activity. J. Biol. Chem. 255, 4834-4842. 

Typical biological compounds that are separated by HIC include, cytochrome P-450, enzymes, DNA polymerase, epidermal growth factor, glycoprotein hormones, human immunoglobulins, human recombinant DNA and canine pancreatic juice proteins. Many HIC techniques have been used for large scale purification of proteins. 

Hsuan, J. Yaish, P. Affinity purification of active epidermal growth factor receptor using monoclonal antibodies. Methods Enzymol., 200, 378-388 (1991)... 

Schulz B, Frommer WB (2004) A plant ABC trans-pcater takes the lotus seat Science 306 622-625 Schulz M, Weissenbock G (1986) Isolation and separation of epidermal and mesophyll protoplasts from rye primary leaves - tissue specific characteristics of secondary phenolic product accumulation. Z Naturforsch [C] 41 22-27 Schumacher HM, Zenk MH (1988) Partial purification and characterization of dihydrobenzophen-anthridine oxidase fmmEschscholtzia californica cell suspension cultures. Plant Cell Rep 7 43 6 Sharma V, Strack D (1985) Vacuolar localization of 1-sinapolglucose 1-malate sinapoyltransferase in protoplasts from cotyledons oiRaphanus sati-vus. Planta 163 563-568... 

We have developed an isolation and purification protocol for STE. The purification has been difficult both due to the cross activity of OTC with 18 0-ACP and the low abundance of STE in epidermal tissues. In sharp contrast to OTE, the purified STE showed high substrate specificity with 18 0-ACP. STE essentially resides in epidermal cells suggesting its role may related to a specific function of epidermal cells. Since wax biosynthesis uses only saturated fatty acids as substrate, the fatty acid (18 0) produced by the hydrolysis of 18 0-ACP by STE can serve as the substrate. Therefore, the role of STE may be to terminate fatty acid biosynthesis in the plastid and supply saturated fatty acids for wax biosynthesis. 

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