Mutagenesis of the crystal contact of acidic fibroblast growth factor

Several mutations at Glu81 located on the crystal contact of human acidic fibroblast growth factor were studied in an effort to improve crystal growth. Mutation to Ser and Thr resulted in crystallization of a rather bulky form of the wild type, whereas mutation to Val prohibited crystallization. These results suggest that crystal growth may be controlled by designing a new interface by protein engineering

Beat-frequency-resolved two-dimensional electronic spectroscopy: disentangling vibrational coherences in artificial fluorescent proteins with sub-10-fs visible laser pulses

We perform a beat-frequency-resolved analysis for two-dimensional electronic spectroscopy using a high-speed and stable 2D electronic spectrometer and few-cycle visible laser pulses to disentangle the vibrational coherences in an artificial fluorescent protein. We develop a highly stable ultrashort light source that generates 5.3-fs visible pulses with a pulse energy of 4.7 uJ at a repetition rate of 10 kHz using multi-plate pulse compression and laser filamentation in a gas cell. The above-5.3-fs laser pulses together with a high-speed multichannel detector enable us to measure a series of 2D electronic spectra, which are resolved in terms of beat frequency related to vibrational coherence. We successfully extract the discrete vibrational peaks behind the inhomogeneous broadening in the absorption spectra and the vibrational quantum beats of the excited electronic state behind the strong stationary signal in the typical 2D electronic spectra

ダイズ グリシニン ノ ケッショウ コウゾウ カイセキ ト カコウ トクセイ ノ カイリョウ ニ カンスル ケンキュウ

京都大学0048新制・論文博士博士(農学)乙第11380号論農博第2513号新制||農||886(附属図書館)学位論文||H16||N3880(農学部図書室)UT51-2004-C128(主査)教授 内海 成, 教授 森 友彦, 教授 廣瀬 正明学位規則第4条第2項該当Doctor of Agricultural ScienceKyoto UniversityDA

X-Ray diffraction experiments for drug target protein of human casein kinase II

Casein kinase II (CKII) exhibits broad phosphorylation reaction on various important regulatory proteins such as survival factors in eukaryotic cells. Since the relationship of CKII over-expression to carcinogenesis and cancer metastasis has been reported, CKII is considered to be one of the drug target proteins. Here, we aimed to obtain the structural information with location of hydration water molecules, and to elucidate the catalytic reaction of CKII for development of effective inhibitors

Fe-S cluster binding mechanism of pigeon\u27s ISCA1 clarified by SEC-SAXS

The iron-sulfur cluster assembly 1 homolog clISCA1 of Columba livia (pigeon) interacts with a quantum biosensor protein clCRY4. The clCRY4/clISCA1 complex tends to orientate along the weak external magnetic-field lines (0.4–10G) under blue light, suggesting that clISCA1 might assist the function of clCRY4. It was expected that the Fe-S cluster binging to clISCA1 might improve the magnetic property of clISCA1. In order to elucidate the Fe-S cluster binging mechanism of clISCA1, we conducted the small angle X-ray scattering analysis coupled with size exclusion chromatography analysis (SEC-SAXS) and UV/Vis spectroscopy. The result suggests that the globular protomers of clISCA1 form columnar oligomers, and Fe-S cluster binding sites are formed between clISCA1 protomers in a columnar oligomer. Periodic and regular binding of Fe-S clusters along the long axis of the columnar oligomer may improve the magnetic susceptibility and the magnetic anisotropy of the clISCA1 oligomer and the clCRY4/clISCA1 complex