Purification and characterization of a novel protamine kinase in HL60 cells

被引:2
作者
Soh, Y [1 ]
Wooten, MW [1 ]
机构
[1] Auburn Univ, Dept Biol Sci, Auburn, AL 36849 USA
关键词
differentiation; HL60; cells; kinase; protamine; protein kinase C;
D O I
10.1093/oxfordjournals.jbchem.a022589
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A protamine kinase from HL60 cells was purified to near homogeneity by DEAE-Sephacel, protamine-agarose, Hydroxylapatite, and S-200 chromatography. It was purified by 75.8-fold through four chromatographic steps, and 0.67% of total activity was recovered. The purified enzyme had an apparent molecular mass of 120 kDa and was activated by Mg2+ or Mn2+, but inhibited by Ca2+, Neither phospholipid nor phorbol ester significantly affected the enzyme activity. Staurosporine was the most potent inhibitor of the enzyme among the protein kinase inhibitors tested, K-252a, H-7, heparin, and staurosporine. The purified protamine kinase exhibited a maximum velocity of 5,000 pmol/min/mg and K-m of 1.3 mM for protamine sulfate as a substrate. Myelin basic protein and protamine sulfate served as the best substrates for the protamine kinase among those tested. The activity of the protamine kinase remained unchanged upon treatment with PMA, retinoic acid, dimethyl sulfoxide, or 1,25 dihydroxy vitamin D-3 for 15 min, while treatment with a differentiating agent, 1,25 dihydroxy vitamin D-3, for one week increased its activity. These results suggest that protamine kinase in HL60 cells is involved in the late stage of the macrophage-monocytic differentiation pathway and may play a role in maintenance of the differentiation after HL60 cells are committed.
引用
收藏
页码:95 / 103
页数:9
相关论文
共 30 条
[1]   MAD - A HETERODIMERIC PARTNER FOR MAX THAT ANTAGONIZES MYC TRANSCRIPTIONAL ACTIVITY [J].
AYER, DE ;
KRETZNER, L ;
EISENMAN, RN .
CELL, 1993, 72 (02) :211-222
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]   INDUCTION OF DIFFERENTIATION OF THE HUMAN PROMYELOCYTIC LEUKEMIA-CELL LINE (HL-60) BY RETINOIC ACID [J].
BREITMAN, TR ;
SELONICK, SE ;
COLLINS, SJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (05) :2936-2940
[4]  
Candeliere GA, 1996, MOL CELL BIOL, V16, P584
[5]   PROTAMINE INDUCES AUTOPHOSPHORYLATION OF PROTEIN-KINASE-C - STIMULATION OF PROTEIN KINASE-C-MEDIATED PROTAMINE PHOSPHORYLATION BY HISTONE [J].
CHAUHAN, VPS ;
CHAUHAN, A .
LIFE SCIENCES, 1992, 51 (07) :537-544
[6]   TERMINAL DIFFERENTIATION OF HUMAN PROMYELOCYTIC LEUKEMIA-CELLS INDUCED BY DIMETHYL-SULFOXIDE AND OTHER POLAR COMPOUNDS [J].
COLLINS, SJ ;
RUSCETTI, FW ;
GALLAGHER, RE ;
GALLO, RC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1978, 75 (05) :2458-2462
[7]   PURIFICATION AND PROPERTIES OF A PROTAMINE KINASE AND A TYPE-II CASEIN KINASE FROM BOVINE KIDNEY MITOCHONDRIA [J].
DAMUNI, Z ;
REED, LJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1988, 262 (02) :574-584
[8]  
DAMUNI Z, 1989, J BIOL CHEM, V264, P6412
[9]   A NOVEL PROTAMINE KINASE-ACTIVITY IN HUMAN PROMYELOCYTIC LEUKEMIA-CELLS [J].
DURHAM, JP ;
BUTCHER, FR ;
FONTANA, JA .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1982, 108 (02) :840-845
[10]   MICROTUBULE-ASSOCIATED-PROTEIN (MAP) KINASE ACTIVATED BY NERVE GROWTH-FACTOR AND EPIDERMAL GROWTH-FACTOR IN PC12 CELLS - IDENTITY WITH THE MITOGEN-ACTIVATED MAP KINASE OF FIBROBLASTIC CELLS [J].
GOTOH, Y ;
NISHIDA, E ;
YAMASHITA, T ;
HOSHI, M ;
KAWAKAMI, M ;
SAKAI, H .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 193 (03) :661-669