Purification and some properties of a latent form cathepsin L from mackerel white muscle

被引:12
作者
Aoki, T [1 ]
Nakano, T [1 ]
Ueno, R [1 ]
机构
[1] MIE UNIV,FAC BIORESOURCES,TSU,MIE 514,JAPAN
关键词
cysteine protease; cathepsin L; mackerel muscle; purification;
D O I
10.2331/fishsci.63.824
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
A latent form cysteine protease was purified from the white muscle of common mackerel Scomber japonicus by ammonium sulfate fractionation and successive column chromatography. The final preparation appeared to be homogeneous on polyacrylamide gel electrophoresis. The molecular mass of the latent form was estimated to be 65 kDa by gel filtration and 69 kDa by SDS-PAGE. The latent form enzyme was activated by acid treatment or pepsin treatment, and hydrolyzed Z-Phe-Arg-MCA, hemoglobin and azocasein. The active form enzyme after acid treatment was enhanced by sulfhydryl compounds, such as dithiothreitol and cysteine, strongly inactivated by leupeptin and E-64. Consequently, we concluded that the active form cysteine protease corresponded to cathepsin L.
引用
收藏
页码:824 / 829
页数:6
相关论文
共 27 条
[1]   PURIFICATION AND CHARACTERIZATION OF A CATHEPSIN B-LIKE ENZYME FROM MACKEREL WHITE MUSCLE [J].
AOKI, T ;
YAMASHITA, T ;
UENO, R .
FISHERIES SCIENCE, 1995, 61 (01) :121-126
[2]   HUMAN-KIDNEY CATHEPSIN-B AND CATHEPSIN-L - CHARACTERIZATION AND POTENTIAL ROLE IN DEGRADATION OF GLOMERULAR BASEMENT-MEMBRANE [J].
BARICOS, WH ;
ZHOU, YW ;
MASON, RW ;
BARRETT, AJ .
BIOCHEMICAL JOURNAL, 1988, 252 (01) :301-304
[3]  
BARRETT AJ, 1981, METHOD ENZYMOL, V80, P535
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   ISOLATION AND SEQUENCE OF A CDNA FOR HUMAN PRO-(CATHEPSIN-L) [J].
GAL, S ;
GOTTESMAN, MM .
BIOCHEMICAL JOURNAL, 1988, 253 (01) :303-306
[7]   EFFECT OF PROTEINASE-INHIBITORS ON INTRACELLULAR PROCESSING OF CATHEPSIN-B, CATHEPSIN-H AND CATHEPSIN-L IN RAT MACROPHAGES [J].
HARA, K ;
KOMINAMI, E ;
KATUNUMA, N .
FEBS LETTERS, 1988, 231 (01) :229-231
[8]   CATHEPSIN-L - NEW PROTEINASE FROM RAT-LIVER LYSOSOMES [J].
KIRSCHKE, H ;
LANGNER, J ;
WIEDERANDERS, B ;
ANSORGE, S ;
BOHLEY, P .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1977, 74 (02) :293-301
[9]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[10]  
MACH L, 1994, J BIOL CHEM, V269, P13030