Amino acids in the N-terminal region regulate the photocycle of photoactive yellow protein
Amino acids in the N-terminal region regulate the photocycle of photoactive yellow protein
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DOI：
10.1093/oxfordjournals.jbchem.a002961
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发表时间：
2001-07-01
影响因子：
2.7
通讯作者：
Kataoka, M
中科院分区：
文献类型：
--
作者：
Harigai, M;Yasuda, S;Kataoka, M
研究了凝乳酶部分消化光活性黄蛋白(PYP)的光谱性质。经DEAE-Sepharose柱层析、蛋白质测序和质谱分析表明，胰凝乳酶能裂解N端的6、15和23个氨基酸(T6、T15和T23)，吸收峰蓝移；T6、T15和T23产生的近紫外中间体的吸收光谱与完整的PYP相同，但寿命分别延长了140、2、300和4,500倍。这些观察结果表明，PYP从近紫外光中间体恢复暗状态的过程被N-末端区域加速，并且该区域对PYP的光循环起着调节作用。
The spectroscopic properties of photoactive yellow protein (PYP) partially digested by chymotrypsin were studied. Chymotrypsin yielded three major products that were yellow but distinguishable by SDS-PAGE, They were readily separated by DEAE-Sepharose column chromatography, Protein sequencing and mass spectrometry demonstrated that chymotrypsin cleaved the N-terminal 6, 15, or 23 amino acids (T6, T15, and T23), The blue-shifts of the absorption ma;bma and the increases in the apparent pK(a) of the chromophores relative to those of intact PYP were less than 4 nm and 0,2, respectively, The absorption spectra of the near-UV intermediates produced from T6, T15, and T23 were identical to that of intact PYP, but with lifetimes that were 140, 2,300, and 4,500 times longer, respectively. These observations suggest that the recovery of the dark state of PYP from the near-UV intermediate is accelerated by the N-terminal region, and that this region acts as a regulatory factor for the photocycle of PYP.