Abstract
A cysteine proteinase inhibitor was found in human nail extract treated with 0.01 M Tris HCl buffer, pH 8.0. It had a 2-fold lower and a 4.5-fold higher activity than that of human skin and hair extracts, respectively. From 5.9 g of human nail, 0.1 mg of cysteine proteinase inhibitor was obtained. It was purified by sequential DE-52 ion exchange and carboxymethyl papain affinity chromotography. The purified inhibitor had an apparent molecular mass of 12 kDa as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. It was more stable against heat and pH than most other proteins. Immunologically, it had the same antigenicity compared with human epidermal cystatin A. Its N-terminal amino acid sequence showed a mixed form comprising a full-length MIPG sequence a truncated IPGG sequence. This sequence was identical to human cystatin A consisting of 20% of the full-length and 80% of the truncated form. These results showed that human nail also contains cystatin A type cysteine proteinase inhibitor. Nails can be used as a source of cystatin A.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Anastasi A, Brown MA, Kembhavi AA, Nicklin MJH, Sayers CA, Sunter DC, Barrett AJ (1983) Cystatin, a protein inhibitor of cysteine proteinases. Biochem J 211: 129–138
Barrett AJ, Kirschke H (1981) Cathepsin B, H, and L. Methods Enzymol 80: 535–563
Barrett AJ, Rawling ND, Davies ME, Machleidt W, Salvesen G, Turk V (1986) Cysteine proteinase inhibitors of the cystatin superfamily. In: Barrett AJ, Salvesen G (eds) Proteinase inhibitors. Elsevier, Amserdam, pp 515–569
Barrett AJ, Fritz H, Grubb A, Isemura S, JÄrvinen M, Katunuma N, Machliedt W, Müller-Esterl W, Sasaki M, Turk V (1986) Nomenclature and classification of the proteins homologous with the cysteine-proteinase inhibitor chicken cystatin. Biochem J 236: 311–312
Björck L, Akesson P, Bohus M, Trojnar J, Abrahamson M, Olafsson T, Grubb A (1989) Bacterial growth blocked by a synthetic peptide based on the structure of a human proteinase inhibitor. Nature 337: 385–386
Björck L, Grubb A, Kjellen L (1990) Cystatin C, a human proteinase inhibitor, blocks replication of herpes simplex virus. J Virol 64: 941–943
Dawber RBR, Baran R (1992) Disorders of nails. In: Champion RH, Burton JL, Ebling FTG (eds) Textbook of dermatology, vol 5. Blackwell Scientific, Oxford, pp 2497–2532
Hawley-Nelson P, Roop DR, Cheng CK, Krieg TM, Yuspa SH (1989) Molecular cloning of mouse epidermal cystatin A and detection of regulated expression in differentiation and tumorigenesis. Mol Carcinogen 1: 202–211
Hewick RM, Hunkapilla MW, Hood LE, Dreyer WJ (1981) A gas-liquid solid phase peptide and protein sequenator. J Biol Chem 256: 7990–7997
Hopsu-Havu VK, Joronen IA, JÄrvinen M, Rinne A (1983) Cysteine proteinase inhibitors in psoriatic epidermis. Arch Dermatol Res 275: 305–309
Hopsu-Havu VK, Joronen I, Rinne A, JÄrvinen M (1987) The role of cysteine proteinase inhibitors in the control of epidermal cell proliferation. J Invest Dermatol 89: 313
JÄrvinen M (1978) Purification and some characteristics of the human epidermal SH-protease inhibitor. J Invest Dermatol 71: 114–118
JÄrvinen M, Hopsu-Havu (1975) α-N-Benzoylarginine-2-naphthylamide hydrolase (cathepsin B1?) from rat skin. II. Purification of the enzyme and demonstration of two inhibitors in the skin. Acta Chem Scand [B] 29: 772–780
JÄrvinen M, Rinne A (1982) Human spleen cysteine proteinase inhibitor. Purification, fractionation into isoelectric variants and some properties of the variants. Biochim Biophys Acta 708: 210–217
Korant BD, Brzin J, Turk V (1985) Cystatin, a protein inhibitor of cysteine proteases alters viral protein cleavages in infected human cells. Biochem Biophys Res Commun 127: 1072–1076
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Lowry OH, Rosebrough NJ, Farr AJ, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275
Rinne A, Kirschke H, JÄrvinen M, Hopsu-Havu VK, Wiederanders B, Bohley P (1985) Localization of cathepsin H and its inhibitor in the skin and other stratified epithelia. Arch Dermatol Res 277: 190–194
Takeda A, Kaji H, Nakamura Y, Samejima T (1989) Comparative studies on the primary structures of human cystatin As from epidermis, liver, spleen, and leukocytes. J Biochem 105: 986–991
Towbin H, Stehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 83: 4350–4354
Tsushima H, Ueki A, Mine H, Nakajima Y, Sumi H, Hopsu-Havu VK (1992) Purification and characterization of acystatintype cysteine proteinase inhibitor in the human hair shaft. Arch Dermatol Res 284: 380–385
Turk V, Bode W (1991) The cystatins: protein inhibitors of cysteine proteinases. FEBS Lett 285: 213–219
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tsushima, H. Isolation of cysteine proteinase inhibitor, cystatin A, from human nails. Arch Dermatol Res 285, 418–422 (1993). https://doi.org/10.1007/BF00372136
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00372136