Chen Y.-LChang P.-JHuang C.-T.CHING-TSAN HUANG2021-07-262021-07-2620181757598https://www.scopus.com/inward/record.uri?eid=2-s2.0-85052952647&doi=10.1007%2fs00253-018-9331-8&partnerID=40&md5=c6f75f00eaa86b1ae0bc36b37f7f7316https://scholars.lib.ntu.edu.tw/handle/123456789/573314The protrusion (P) domain of the major structural protein VP1 of norovirus (NoV) is critical for the host’s immune response and receptor binding. Most heterologous P domains expressed in Escherichia coli or Komagataella pastoris (formally known as Pichia pastoris) form P particles consisting of 24 P monomers formed through intermolecular contact in the P regions and an end-linked cysteine tag. The small P particle is only found in P domains with terminal modifications. In this study, the NoV P domain of the most predominant NoV strain GII.4 isolated from Taiwan was expressed in K. pastoris. A high yield of NoV P was obtained using the high-cell density fermentation process in K. pastoris. A large amount of the small P particles and the trimer and dimer complexes formed by 12, 6, and 2 P monomers were observed in both the expression of the NoV P-His and P containing cysteine tag at the N-terminus. Dynamic light scattering and transmission electron microscopy analysis of the purified NoV P-His and P revealed that most of these small P particles are triangle-, square-, and ring-shaped with a diameter of 14–15?nm. The binding ability of purified NoV P-His and P to human histo-blood group antigen was confirmed by a saliva-binding assay. Without terminal modification, small P particles were formed in our study. The amino acid sequence analysis showed only four different amino acids (residue 84, 119, 136, and 313) between the P domain in this study and other investigated GII.4 strains suggesting that these amino acids might play an important role in the P particle formation. The small P particles formed by the Taiwan-native norovirus P domain overexpressed in K. pastoris may provide further information for morphogenesis studies and vaccine development. ? 2018, Springer-Verlag GmbH Germany, part of Springer Nature.Dimers; Escherichia coli; High resolution transmission electron microscopy; Light scattering; Light transmission; Monomers; Purification; Strain; Transmission electron microscopy; Yeast; Amino acid sequence analysis; Blood group antigen; High cell density fermentations; Intermolecular contacts; Norovirus; Pichia Pastoris; Terminal modifications; Transmission electron; Amino acids; cysteine; dimer; monomer; protein VP1; structural protein; viral protein; antigen; chemical binding; coliform bacterium; fermentation; gene expression; immune response; particulate matter; protein; virus; amino acid sequence; amino terminal sequence; Article; enzyme linked immunosorbent assay; fermentation; Komagataella pastoris; liquid chromatography-mass spectrometry; nonhuman; Norovirus; particle size; photon correlation spectroscopy; polyacrylamide gel electrophoresis; protein expression; saliva; transmission electron microscopy; virus particle; Western blotting; budding yeast; chemistry; gene expression; genetics; human; metabolism; Norovirus; protein domain; protein motif; Taiwan; Escherichia coli; Norovirus; Pichia pastoris; Amino Acid Motifs; Gene Expression; Humans; Norovirus; Protein Domains; Saccharomycetales; Taiwan; Viral Structural Proteins[SDGs]SDG3Dimers; Escherichia coli; High resolution transmission electron microscopy; Light scattering; Light transmission; Monomers; Purification; Strain; Transmission electron microscopy; Yeast; Amino acid sequence analysis; Blood group antigen; High cell density fermentations; Intermolecular contacts; Norovirus; Pichia Pastoris; Terminal modifications; Transmission electron; Amino acids; cysteine; dimer; monomer; protein VP1; structural protein; viral protein; antigen; chemical binding; coliform bacterium; fermentation; gene expression; immune response; particulate matter; protein; virus; amino acid sequence; amino terminal sequence; Article; enzyme linked immunosorbent assay; fermentation; Komagataella pastoris; liquid chromatography-mass spectrometry; nonhuman; Norovirus; particle size; photon correlation spectroscopy; polyacrylamide gel electrophoresis; protein expression; saliva; transmission electron microscopy; virus particle; Western blotting; budding yeast; chemistry; gene expression; genetics; human; metabolism; Norovirus; protein domain; protein motif; Taiwan; Escherichia coli; Norovirus; Pichia pastoris; Amino Acid Motifs; Gene Expression; Humans; Norovirus; Protein Domains; Saccharomycetales; Taiwan; Viral Structural Proteinsjournal article10.1007/s00253-018-9331-8301871002-s2.0-85052952647