title: Identification and characterization of Laodelphax striatellus (Insecta: Hemiptera: Delphacidae) neutral sphingomyelinase.
abstract: The neutral sphingomyelinase (nSMase) 1 homologue gene LsSMase was cloned from Laodelphax striatellus, a direct sap-sucker and virus vector of gramineous plants, and expressed via a Bac to Bac baculovirus expression system. The LsSMase-enhanced green fluorescent protein fusion protein was located in the endoplasmic reticulum in a similar manner to mammalian nSMase 1. The biochemical properties of LsSMase were determined in detail. The optimal pH and temperature for recombinant LsSMase were 8 and 37 °C, respectively. LsSMase was an Mg2+or Mn2+dependent enzyme, but different concentration of each were needed. The activity of LsSMase was significantly stimulated by Ethylene glycol bis(2-aminoethyl ether)tetraacetic acid (EGTA), whereas it was inhibited by ethylenediaminetetraacetic acid. Millimolar concentrations of Zn2+completely inhibited LsSMase. The reducing agents dithiothreitol and β-mercaptoethanol varied in their effects on activity. Phospholipids were not found to stimulate LsSMase.
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.|Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: X-W
authors.initials: XW
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: M-A
authors.initials: MA
authors.lastname: Begum
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: C-H
authors.initials: CH
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: X-X
authors.initials: XX
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: W-J
authors.initials: WJ
authors.lastname: Jiao
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Y-R
authors.initials: YR
authors.lastname: Zhang
authors.affiliation: Center for Chemical Analysis and Detection, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: J-Q
authors.initials: JQ
authors.lastname: Yuan
authors.affiliation: Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: H-Y
authors.initials: HY
authors.lastname: Li
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, Jiangsu, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Department of Medicine, State University of New York at Stony Brook. Stony Brook, NY, USA.
authors.forename: C
authors.initials: C
authors.lastname: Mao
authors.affiliation: State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Z-R
authors.initials: ZR
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006430:Hemiptera / Q000201:enzymology* / Q000235:genetics; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013108:Sphingomyelin Phosphodiesterase / Q000235:genetics / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D013108:Sphingomyelin Phosphodiesterase
keywords: Laodelphax striatellus; insect; sphingolipid; sphingomyelinase