The clotting protein(CP) plays important and diverse roles in crustaceans,such as coagulation and lipid transportation.A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis(name...The clotting protein(CP) plays important and diverse roles in crustaceans,such as coagulation and lipid transportation.A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis(named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography.Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca2+,suggesting that the clotting reaction is catalyzed by a Ca2+-dependent transglutaminase in shrimp hemocytes.The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE.CP exists as disulfide-linked homodimers and oligomers.The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon,Farfantepenaeus paulensis and Litopenaeus vannamei;and similar to that of other decapods.The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry.Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.展开更多
Proteomic analysis of upland cotton was performed to profile the global detectable proteomes of ovules and fibers using two-dimensional electrophoresis(2DE).A total of 1,203 independent protein spots were collected fr...Proteomic analysis of upland cotton was performed to profile the global detectable proteomes of ovules and fibers using two-dimensional electrophoresis(2DE).A total of 1,203 independent protein spots were collected from representative 2DE gels,which were digested with trypsin and identified by matrix-assisted laser desorption and ionization-time-offlight/time-of-flight(MALDI-TOF/TOF)mass spectrometry.The mass spectrometry or tandem mass spectrometry(MS or MS/MS)data were then searched against a local database constructed from Gossypium hirsutum genome sequences,resulting in successful identification of 975 protein spots(411 for ovules and 564 for fibers).Functional annotation analysis of the 975identified proteins revealed that ovule-specific proteins were mainly enriched in functions related to fatty acid elongation,sulfur amino acid metabolism and post-replication repair,while fiber-specific proteins were enriched in functions related to root hair elongation,galactose metabolism and D-xylose metabolic processes.Further annotation analysis of the most abundant protein spots showed that 28.96%of the total proteins in the ovule were mainly located in the Golgi apparatus,endoplasmic reticulum,mitochondrion and ribosome,whereas in fibers,27.02%of the total proteins were located in the cytoskeleton,nuclear envelope and cell wall.Quantitative real-time polymerase chain reaction(q RT-PCR)analyses of the ovule-specific protein spots P61,P93 and P198 and fiber-specific protein spots 230,477 and 511 were performed to validate the proteomics data.Protein-protein interaction network analyses revealed very different network cluster patterns between ovules and fibers.This work provides the largest protein identification dataset of 2DE-detectable proteins in cotton ovules and fibers and indicates potentially important roles of tissue-specific proteins,thus providing insights into the cotton ovule and fiber proteomes on a global scale.展开更多
基金supported by the National Natural Science Foundation of China(No.30600458)
文摘The clotting protein(CP) plays important and diverse roles in crustaceans,such as coagulation and lipid transportation.A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis(named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography.Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca2+,suggesting that the clotting reaction is catalyzed by a Ca2+-dependent transglutaminase in shrimp hemocytes.The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE.CP exists as disulfide-linked homodimers and oligomers.The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon,Farfantepenaeus paulensis and Litopenaeus vannamei;and similar to that of other decapods.The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry.Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.
基金the Special Fund for Agro-scientific Research in the Public Interest of the People’s Republic of China (201403075)Major Technology Project of Hainan (ZDZX2013010-1)+1 种基金Program for Top Young Talents in the Chinese Academy of Tropical Agricultural Sciences (ITBB130102)China Postdoctoral Science Foundation (20110490003)
文摘Proteomic analysis of upland cotton was performed to profile the global detectable proteomes of ovules and fibers using two-dimensional electrophoresis(2DE).A total of 1,203 independent protein spots were collected from representative 2DE gels,which were digested with trypsin and identified by matrix-assisted laser desorption and ionization-time-offlight/time-of-flight(MALDI-TOF/TOF)mass spectrometry.The mass spectrometry or tandem mass spectrometry(MS or MS/MS)data were then searched against a local database constructed from Gossypium hirsutum genome sequences,resulting in successful identification of 975 protein spots(411 for ovules and 564 for fibers).Functional annotation analysis of the 975identified proteins revealed that ovule-specific proteins were mainly enriched in functions related to fatty acid elongation,sulfur amino acid metabolism and post-replication repair,while fiber-specific proteins were enriched in functions related to root hair elongation,galactose metabolism and D-xylose metabolic processes.Further annotation analysis of the most abundant protein spots showed that 28.96%of the total proteins in the ovule were mainly located in the Golgi apparatus,endoplasmic reticulum,mitochondrion and ribosome,whereas in fibers,27.02%of the total proteins were located in the cytoskeleton,nuclear envelope and cell wall.Quantitative real-time polymerase chain reaction(q RT-PCR)analyses of the ovule-specific protein spots P61,P93 and P198 and fiber-specific protein spots 230,477 and 511 were performed to validate the proteomics data.Protein-protein interaction network analyses revealed very different network cluster patterns between ovules and fibers.This work provides the largest protein identification dataset of 2DE-detectable proteins in cotton ovules and fibers and indicates potentially important roles of tissue-specific proteins,thus providing insights into the cotton ovule and fiber proteomes on a global scale.
