Human skin proteases: separation and characterization of two alkaline proteases, one splitting trypsin and the other chymotrypsin substrates

Archives of Dermatological Research
J E Fräki, V K Hopsu-Havu

Abstract

Two alkaline proteases, one splitting preferentially the substrates of chymotrypsin (ATEE) and the other one those of trypsin (BAEE), were separated and partially purified by chromatographic means from human skin extract made in a buffer containing 1.07 mol/1 KC1. The proteins soluble in dilute buffer were removed by a prior extraction. The enzymes could be separated effectively only in the presence of KC1 at a high conc-ntration since large molecular size aggregates or polymers were formed in solutions of low ionic strength. In the presence of 2 mol/1 KC1 the molecular size of the BAEE-hydrolysing enzyme was 120000 and that of the ATEE-hydrolysing enzyme 30000. The ATEE-hydrolysing enzyme was purified by Sephadex G-100 gel filtration and DEAE-cellulose chromatography about 250 fold. It also hydrolysed esters of tryptophane and phenylalanine as well as casein with optimum pH 7.8--8.2. The enzyme was inhibited effectively by LBTI, SBTI and partially by trasylol, TPCK and TLCK, but not by E-600 and SH-modifers. The hydrolysis of ATEE was doubled in the presence of 1 mol/lKCl, NaCl, KBr or NaBr but that of casein was inhibited to some extent. Human serum and alpha-1-antitrypsin inhibited this enzyme but not C1-inactivator. alpha-2...Continue Reading

References

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Citations

Jan 1, 1989·Archives of Dermatological Research·I T HarvimaJ E Fräki
Aug 1, 1986·The British Journal of Dermatology·R M LavkerG S Lazarus
Mar 31, 1979·Archives of Dermatological Research·J E FräkiV K Hopsu-Havu
Jan 1, 1986·Archives of Dermatological Research·J E FräkiG S Lazarus
Jan 1, 1989·Journal of Periodontal Research·S W Cox, B M Eley
Jul 21, 1977·Archives of Dermatological Research·J E Fräki, V K Hopsu-Havu
Jan 1, 1990·Archives of Dermatological Research·I T HarvimaM Horsmanheimo
Oct 28, 1999·Experimental Dermatology·B AlgermissenB M Henz
Jun 1, 1979·Journal of Cutaneous Pathology·J E FräkiV K Hopsu-Havu
Apr 1, 1981·International Journal of Dermatology·V K Hopsu-Havu, J E Fräki
Jul 1, 1989·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·R C Benyon
Jan 1, 1993·Archives of Dermatological Research·I T HarvimaM Horsmanheimo
Dec 1, 1979·Clinical and Experimental Dermatology·J E Fräki
Jan 1, 1984·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·N HorieW L Epstein
Sep 21, 1988·Biochimica Et Biophysica Acta·I T HarvimaJ E Fräki
Feb 1, 1978·The Journal of Investigative Dermatology·H E Seppä, M Järvinen
Jun 1, 1986·Analytical Biochemistry·L A JohnsonM Eisenberg
Nov 30, 1984·Biochemical and Biophysical Research Communications·M EisenbergK E Moon
Sep 1, 1980·Experimental Eye Research·S S Tse, B J Ortwerth
Oct 1, 1984·Journal of the American Academy of Dermatology·G LipkinF Kueppers

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