CA2+-CALMODULIN-DEPENDENT PROTEIN KINASE-IA AND KINASE-IB FROM RAT-BRAIN .1. IDENTIFICATION, PURIFICATION, AND STRUCTURAL COMPARISONS

被引:0
|
作者
DEREMER, MF
SAELI, RJ
EDELMAN, AM
机构
[1] SUNY BUFFALO, DEPT PHARMACOL & THERAPEUT, 102 FARBER HALL, BUFFALO, NY 14260 USA
[2] SUNY BUFFALO, DEPT BIOL SCI, BUFFALO, NY 14260 USA
关键词
D O I
暂无
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
Two Ca2+-calmodulin (CaM)-dependent protein kinases were purified from rat brain using as substrate a synthetic peptide based on site 1 (site 1 peptide) of the synaptic vesicle-associated protein, synapsin I. One of the purified enzymes was an approximately 89% pure protein of M(r) = 43,000 which bound CaM in a Ca2+-dependent fashion. The other purified enzyme was an apparently homogenous protein of M(r) = 39,000 accompanied by a small amount of a M(r) = 37,000 form which may represent a proteolytic product of the 39-kDa enzyme. The 39-kDa protein bound CaM in a Ca2+-dependent fashion. Gel filtration analysis indicated that both enzymes are monomers. The 43- and 39-kDa enzymes are named Ca2+-CaM-dependent protein kinases Ia and Ib (CaM kinases Ia, Ib), respectively. The specific activities of CaM kinases Ia and Ib were similar (5-8-mu-mol/min/mg protein). CaM kinase Ia (but not CaM kinase Ib) activity was enhanced by addition of a CaM-Sepharose column wash (non-binding) fraction suggesting the existence of an "activator" of CaM kinase la. Both kinases phosphorylated exogenous substrates (site 1 peptide and synapsin I) in a Ca2+-CaM-dependent fashion and both kinases underwent autophosphorylation. CaM kinase Ia autophosphorylation was Ca2+-CaM-dependent and occurred exclusively on threonine while CaM kinase Ib autophosphorylation showed Ca2+-CaM independence and occurred on both serine and threonine. Proteolytic digestion of autophosphorylated CaM kinases Ia and Ib yielded phosphopeptides of differing M(r). These characteristics, as well as enzymatic and regulatory properties (DeRemer, M. F., Saeli, R. J. Brautigen, D. L., and Edelman, A. M. (1992) J. Biol. Chem. 267, 13466-13471), indicate that CaM kinases Ia and Ib are distinct and possibly previously unrecognized enzymes.
引用
收藏
页码:13460 / 13465
页数:6
相关论文
共 50 条
  • [41] A NOVEL PROTEIN-ACTIVATION OF CA2+ CALMODULIN-DEPENDENT PROTEIN KINASE-I PURIFIED AND CHARACTERIZED FROM RAT-BRAIN
    NAITO, Y
    INOUE, S
    MIZUTANI, A
    SUGITA, R
    YOKOKURA, H
    SUDO, T
    HIDAKA, H
    JOURNAL OF NEUROCHEMISTRY, 1995, 65 : S24 - S24
  • [42] DEVELOPMENTAL AND REGIONAL EXPRESSION OF MULTIFUNCTIONAL CA2+/CALMODULIN-DEPENDENT PROTEIN-KINASE ISOFORMS IN RAT-BRAIN
    BROCKE, L
    SRINIVASAN, M
    SCHULMAN, H
    JOURNAL OF NEUROSCIENCE, 1995, 15 (10): : 6797 - 6808
  • [43] REGIONAL DISTRIBUTION OF TYPE-II CA-2+ CALMODULIN-DEPENDENT PROTEIN-KINASE IN RAT-BRAIN
    ERONDU, NE
    KENNEDY, MB
    JOURNAL OF NEUROSCIENCE, 1985, 5 (12): : 3270 - 3277
  • [44] CA2+/CALMODULIN-DEPENDENT PROTEIN-KINASE-V - TISSUE DISTRIBUTION AND IMMUNOHISTOCHEMICAL LOCALIZATION IN RAT-BRAIN
    ITO, T
    MOCHIZUKI, H
    KATO, M
    NIMURA, Y
    HANAI, T
    USUDA, N
    HIDAKA, H
    ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1994, 312 (01) : 278 - 284
  • [45] AKAP79 selectively enhances protein kinase C regulation of GluR1 at a Ca2+-calmodulin-dependent protein kinase II/protein kinase C site
    Tavalin, Steven J.
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2008, 283 (17) : 11445 - 11452
  • [46] IMMUNOHISTOCHEMICAL LOCALIZATION OF CA-2+/CALMODULIN-DEPENDENT PROTEIN KINASE-II IN RAT-BRAIN AND VARIOUS TISSUES
    FUKUNAGA, K
    GOTO, S
    MIYAMOTO, E
    JOURNAL OF NEUROCHEMISTRY, 1988, 51 (04) : 1070 - 1078
  • [47] Identification and purification of Ca2+/calmodulin-dependent protein kinase V from human gastric carcinoma
    Ohta, H
    Niki, I
    Ito, T
    Kato, M
    Nimura, Y
    Usuda, N
    Hidaka, H
    BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE, 1996, 1317 (03): : 175 - 182
  • [48] CA2+/CALMODULIN-DEPENDENT PROTEIN KINASE-II FROM HEN BRAIN - PURIFICATION AND CHARACTERIZATION
    GUPTA, RP
    LAPADULA, DM
    ABOUDONIA, MB
    BIOCHEMICAL PHARMACOLOGY, 1992, 43 (09) : 1975 - 1988
  • [49] PURIFICATION AND CHARACTERIZATION OF CA-2+ CALMODULIN-DEPENDENT PROTEIN KINASE-I FROM BOVINE BRAIN
    NAIRN, AC
    GREENGARD, P
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1987, 262 (15) : 7273 - 7281
  • [50] PURIFICATION AND CHARACTERIZATION OF CA2+ CALMODULIN-DEPENDENT PROTEIN KINASE-V FROM RAT CEREBRUM
    MOCHIZUKI, H
    ITO, T
    HIDAKA, H
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1993, 268 (12) : 9143 - 9147