caa a22 4500 397494416 CHVBK 20180308164517.0 cr unu---uuuuu 161202e199506 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a124842 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a124842 Characterization of Vitelline Membrane Outer Layer Protein I, VMO-I: Amino Acid Sequence and Structural Stability [Elektronische Daten] [Shoko Kido, Yukio Doi, Fimi Kim, Emi Morishita, Hiroshi Narita, Shigenori Kanaya, Tadayashu Ohkubo, Ken Nishikawa, Toru Yao, Tatsuo Ooi] Vitelline membrane outer layer protein I (VMO-I) tightly bound to ovomucin fibrils of hen's egg yolk membrane was characterized in terms of its amino acid sequence and structural stability. The deduced sequence of VMO-I using the conventional sequencing method is: RTREYTSVITVPNGGHWGKWGIRQFCHSGYANGFALKVEPSQFGRDDTALNGIRLRCLD- GSVIESLVGKWGTWTSFLVCPTGYLVSFSLRSEKSQGGGDDTAANNIQFRCSDEAVLVGD- DLSWGREGPWSKRCKICGLQTKVESPOGLRDDTALNNVRFFCCK. Thus, VMO-I is composed of 163 amino acid residues with a calculated molecular weight of 17,979. The sequence confirms the cDNA sequence of VMO-I we recently determined and does not show any significant similarity to proteins compiled in the NBRF database. Two of the four disulfide bonds found in VMO-I were estimated to lie between Cys26 and Cys57 and between Cys79 and CysllO. The sequence analyses show that VMO-I contains three 53-residue internal repeats that contain distinctive regions of turns flanked by β-sheets consistent with the recent finding that the molecule contains a new β-fold motif, the β-prism. The molecular characteristics of VMO-I in solution were examined by CD spectroscopy in the far and near ultraviolet regions, NMR spectroscopy, and high sensitive differential scanning calor-imetry (DSC). CD spectra in the far UV region at room temperature were similar to that assigned to a random coil, while in the near UV region, small positive peaks were observed. The ellipticity in both regions decreased on raising the temperature. Proton NMR experiments showed the native structure unfolds to unordered conformations at 70°C. A single positive peak appeared in the DSC curve with the transition temperature of 67.5°C, from which an unfolding enthalpy of 107 kcal/mol and a heat capacity change of 2.6 kcal/mol/K were obtained. These results showed a normal degree of stability for the β-prism structure found in VMO-I. © Oxford University Press Regular Papers nationallicence amino acid sequence nationallicence CD nationallicence sequence analysis nationallicence thermal stability nationallicence VMO-I nationallicence Kido Shoko Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut Doi Yukio Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut Kim Fimi Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut Morishita Emi Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut Narita Hiroshi Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut Kanaya Shigenori Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut Ohkubo Tadayashu Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut Nishikawa Ken Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut Yao Toru Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut Ooi Tatsuo Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut The Journal of Biochemistry Oxford University Press 117/6(1995-06), 1183-1191 0021-924X 117:6<1183 1995 117 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a124842 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a124842 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kido Shoko Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut NATIONALLICENCE 700 1- Doi Yukio Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut NATIONALLICENCE 700 1- Kim Fimi Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut NATIONALLICENCE 700 1- Morishita Emi Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut NATIONALLICENCE 700 1- Narita Hiroshi Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut NATIONALLICENCE 700 1- Kanaya Shigenori Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Ohkubo Tadayashu Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Nishikawa Ken Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Yao Toru Protein Engineering Research Institute Furuedai, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Ooi Tatsuo Department of Food Science, Kyoto Women's University Higashiyama-ku, Kyoto 605 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 117/6(1995-06), 1183-1191 0021-924X 117:6<1183 1995 117 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford