TY - JOUR
T1 - SDS-induced oligomerization of Lys49-phospholipase A 2 from snake venom
AU - Matsui, Takashi
AU - Kamata, Shizuka
AU - Ishii, Kentaro
AU - Maruno, Takahiro
AU - Ghanem, Nouran
AU - Uchiyama, Susumu
AU - Kato, Koichi
AU - Suzuki, Atsuo
AU - Oda-Ueda, Naoko
AU - Ogawa, Tomohisa
AU - Tanaka, Yoshikazu
N1 - Funding Information:
The X-ray diffraction experiments were performed at the Photon Factory (proposals 16G092, 18G060, and 17G595) and SPring-8 (proposals 2015B6524, 2016A2565 and 2016B2565). This work was supported in part by the following sources: the Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (to T.Matsui and Y.T.), the PRESTO (to Y.T.), the Joint Research by Exploratory Research Center on Life and Living Systems (ExCELLS) (to SU and KK), and the Platform Project for Supporting Drug Discovery and Life Science Research [Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)] from the Japan Agency for Medical Research and Development (AMED) under Grant Number JP18am0101095 (to T.Matsui). This research was also supported by a JSPS Grants-in-Aid for Scientific Research on Innovative Areas entitled “Dynamical Ordering of Biomolecular Systems for Creation of Integrated Functions” (25102008, 16H00770, and 16H00748), NEDO, and JICE.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Phospholipase A 2 (PLA 2 ) is one of the representative toxic components of snake venom. PLA 2 s are categorized into several subgroups according to the amino acid at position 49, which comprises either Asp49, Lys49, Arg49 or Ser49. Previous studies suggested that the Lys49-PLA 2 assembles into an extremely stable dimer. Although the behavior on Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing or non-reducing conditions suggested the presence of intermolecular disulfide bonds, these bonds were not observed in the crystal structure of Lys49-PLA 2 . The reason for this discrepancy between the crystal structure and SDS-PAGE of Lys49-PLA 2 remains unknown. In this study, we analyzed a Lys49-PLA 2 homologue from Protobothrops flavoviridis (PflLys49-PLA 2 BPII), by biophysical analyses including X-ray crystallography, SDS-PAGE, native-mass spectrometry, and analytical ultracentrifugation. The results demonstrated that PflLys49-PLA 2 BPII spontaneously oligomerized in the presence of SDS, which is one of the strongest protein denaturants.
AB - Phospholipase A 2 (PLA 2 ) is one of the representative toxic components of snake venom. PLA 2 s are categorized into several subgroups according to the amino acid at position 49, which comprises either Asp49, Lys49, Arg49 or Ser49. Previous studies suggested that the Lys49-PLA 2 assembles into an extremely stable dimer. Although the behavior on Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing or non-reducing conditions suggested the presence of intermolecular disulfide bonds, these bonds were not observed in the crystal structure of Lys49-PLA 2 . The reason for this discrepancy between the crystal structure and SDS-PAGE of Lys49-PLA 2 remains unknown. In this study, we analyzed a Lys49-PLA 2 homologue from Protobothrops flavoviridis (PflLys49-PLA 2 BPII), by biophysical analyses including X-ray crystallography, SDS-PAGE, native-mass spectrometry, and analytical ultracentrifugation. The results demonstrated that PflLys49-PLA 2 BPII spontaneously oligomerized in the presence of SDS, which is one of the strongest protein denaturants.
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U2 - 10.1038/s41598-019-38861-8
DO - 10.1038/s41598-019-38861-8
M3 - Article
C2 - 30787342
AN - SCOPUS:85061784759
VL - 9
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 2330
ER -