Nevalainen L T, Aoyama T, Ikura M, Crivici A, Yan H, Chua N H, Nairn A C
Laboratory of Plant Molecular Biology, Rocketeller University, New York, NY 10021, USA.
Biochem J. 1997 Jan 1;321 ( Pt 1)(Pt 1):107-15. doi: 10.1042/bj3210107.
We describe the isolation and interaction with calmodulin (CaM) of two 10-amino-acid peptides (termed peptides 1 and 2; AWDTVRISFG and AWPSLQAIRG respectively) derived from a phage random peptide display library. Both peptides are shorter than previously described CaM-binding peptides and lack certain features found in the sequences of CaM-binding domains present in CaM-activated enzymes. However, 1H NMR spectroscopy and fluorimetry indicate that both peptides interact with CaM in the presence of Ca2+. The two peptides differentially inhibited CaM-dependent kinases I and II (CaM kinases I and II) but did not affect CaM-dependent phosphodiesterase. Peptide 1 inhibited CaM kinase I but not CaM kinase II, whereas peptide 2 inhibited CaM kinase II, but only partially inhibited CaM kinase I at a more than 10-fold higher concentration. Peptide 1 also inhibited a plant calcium-dependent protein kinase, whereas peptide 2 did not. The ability of peptides 1 and 2 to differentially inhibit CaM-dependent kinases and CaM-dependent phosphodiesterase suggests that they may bind to distinct regions of CaM that are specifically responsible for activation of different CaM-dependent enzymes.
我们描述了从噬菌体随机肽展示文库中获得的两个10氨基酸肽（分别称为肽1和肽2；序列为AWDTVRISFG和AWPSLQAIRG）与钙调蛋白（CaM）的分离及相互作用。这两个肽均比先前描述的CaM结合肽短，且缺乏CaM激活酶中存在的CaM结合域序列所具有的某些特征。然而，1H核磁共振光谱和荧光测定表明，在Ca2+存在的情况下，这两个肽均能与CaM相互作用。这两个肽对CaM依赖性激酶I和II（CaM激酶I和II）具有不同程度的抑制作用，但不影响CaM依赖性磷酸二酯酶。肽1抑制CaM激酶I，但不抑制CaM激酶II，而肽2抑制CaM激酶II，但只有在浓度高于10倍以上时才对CaM激酶I有部分抑制作用。肽1还抑制一种植物钙依赖性蛋白激酶，而肽2则无此作用。肽1和肽2对CaM依赖性激酶和CaM依赖性磷酸二酯酶具有不同抑制作用的能力表明，它们可能与CaM的不同区域结合，这些区域专门负责激活不同的CaM依赖性酶。
