通过质谱结合埃德曼降解法和二维聚丙烯酰胺凝胶电泳对黄粉虫幼虫的一种表皮蛋白同工型进行研究。
Isoforms of a cuticular protein from larvae of the meal beetle, Tenebrio molitor, studied by mass spectrometry in combination with Edman degradation and two-dimensional polyacrylamide gel electrophoresis.
作者信息
Haebel S, Jensen C, Andersen S O, Roepstorff P
机构信息
Department of Molecular Biology, Odense University, Denmark.
出版信息
Protein Sci. 1995 Mar;4(3):394-404. doi: 10.1002/pro.5560040306.
Abstract
Simultaneous sequencing, using a combination of mass spectrometry and Edman degradation, of three approximately 15-kDa variants of a cuticular protein extracted from the meal beetle Tenebrio molitor larva is demonstrated. The information obtained by matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) time-course monitoring of enzymatic digests was found essential to identify the differences among the three variants and for alignment of the peptides in the sequence. To determine whether each individual insect larva contains all three protein variants, proteins extracted from single animals were separated by two-dimensional gel electrophoresis, electroeluted from the gel spots, and analyzed by MALDI MS. Molecular weights of the proteins present in each sample could be obtained, and mass spectrometric mapping of the peptides after digestion with trypsin gave additional information. The protein isoforms were found to be allelic variants.
摘要
本文展示了使用质谱法和艾德曼降解法相结合的方法，对从黄粉虫幼虫中提取的一种约15 kDa的表皮蛋白的三种变体进行同步测序。通过基质辅助激光解吸电离质谱（MALDI MS）对酶解产物进行时间进程监测所获得的信息，对于识别这三种变体之间的差异以及肽段在序列中的比对至关重要。为了确定每只昆虫幼虫是否都含有这三种蛋白质变体，从单只动物中提取的蛋白质通过二维凝胶电泳进行分离，从凝胶斑点中电洗脱出来，并通过MALDI MS进行分析。可以获得每个样品中存在的蛋白质的分子量，用胰蛋白酶消化后的肽段的质谱图谱给出了更多信息。发现这些蛋白质异构体是等位基因变体。
相似文献
1
Isoforms of a cuticular protein from larvae of the meal beetle, Tenebrio molitor, studied by mass spectrometry in combination with Edman degradation and two-dimensional polyacrylamide gel electrophoresis.通过质谱结合埃德曼降解法和二维聚丙烯酰胺凝胶电泳对黄粉虫幼虫的一种表皮蛋白同工型进行研究。
Protein Sci. 1995 Mar;4(3):394-404. doi: 10.1002/pro.5560040306.
2
Rapid mass spectrometric identification of proteins from two-dimensional polyacrylamide gels after in gel proteolytic digestion.凝胶内蛋白酶解后二维聚丙烯酰胺凝胶中蛋白质的快速质谱鉴定
Electrophoresis. 1997 Mar-Apr;18(3-4):391-402. doi: 10.1002/elps.1150180313.
3
Identification of two-dimensionally separated human cerebrospinal fluid proteins by N-terminal sequencing, matrix-assisted laser desorption/ionization--mass spectrometry, nanoliquid chromatography-electrospray ionization-time of flight-mass spectrometry, and tandem mass spectrometry.通过N端测序、基质辅助激光解吸/电离-质谱、纳升液相色谱-电喷雾电离-飞行时间质谱以及串联质谱鉴定二维分离的人脑脊液蛋白质
Electrophoresis. 2000 Jun;21(11):2266-83. doi: 10.1002/1522-2683(20000601)21:11<2266::AID-ELPS2266>3.0.CO;2-Z.
4
Investigation of charge variants of rViscumin by two-dimensional gel electrophoresis and mass spectrometry.通过二维凝胶电泳和质谱法对重组相思豆毒素电荷变体的研究。
Electrophoresis. 2001 Aug;22(14):2888-97. doi: 10.1002/1522-2683(200108)22:14<2888::AID-ELPS2888>3.0.CO;2-C.
5
Rapid identification of comigrating gel-isolated proteins by ion trap-mass spectrometry.通过离子阱质谱快速鉴定凝胶分离的共迁移蛋白质。
Electrophoresis. 1998 May;19(6):968-80. doi: 10.1002/elps.1150190612.
6
Comparison of vacuum matrix-assisted laser desorption/ionization (MALDI) and atmospheric pressure MALDI (AP-MALDI) tandem mass spectrometry of 2-dimensional separated and trypsin-digested glomerular proteins for database search derived identification.用于数据库搜索鉴定的二维分离和胰蛋白酶消化的肾小球蛋白质的真空基质辅助激光解吸/电离（MALDI）和大气压MALDI（AP-MALDI）串联质谱的比较
J Proteome Res. 2006 Aug;5(8):1967-78. doi: 10.1021/pr060165s.
7
Coupling capillary high-performance liquid chromatography to matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of peptides via automated microblotting onto membrane substrates.将毛细管高效液相色谱与基质辅助激光解吸/电离质谱联用，并通过自动微量印迹到膜基质上对肽段进行N端测序。
Anal Biochem. 1998 Sep 10;262(2):99-109. doi: 10.1006/abio.1998.2755.
8
Identification of proteins separated by one-dimensional sodium dodecyl sulfate/polyacrylamide gel electrophoresis with matrix-assisted laser desorption/ionization ion trap mass spectrometry; comparison with matrix-assisted laser desorption/ionization time-of-flight mass fingerprinting.用基质辅助激光解吸/电离离子阱质谱法鉴定一维十二烷基硫酸钠/聚丙烯酰胺凝胶电泳分离的蛋白质；与基质辅助激光解吸/电离飞行时间质谱指纹图谱的比较。
Rapid Commun Mass Spectrom. 2003;17(17):1995-2004. doi: 10.1002/rcm.1143.
9
Identification of proteins from two-dimensional gel electrophoresis of human erythroleukemia cells using capillary high performance liquid chromatography/electrospray-ion trap-reflectron time-of-flight mass spectrometry with two-dimensional topographic map analysis of in-gel tryptic digest products.利用毛细管高效液相色谱/电喷雾-离子阱-反射电子飞行时间质谱法结合凝胶内胰蛋白酶消化产物的二维地形图分析，从人红白血病细胞的二维凝胶电泳中鉴定蛋白质。
Rapid Commun Mass Spectrom. 1999;13(19):1907-16. doi: 10.1002/(SICI)1097-0231(19991015)13:19<1907::AID-RCM732>3.0.CO;2-T.
10
Identification of phosphorylated proteins from thrombin-activated human platelets isolated by two-dimensional gel electrophoresis by electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS).通过二维凝胶电泳分离，再利用电喷雾电离串联质谱（ESI-MS/MS）和液相色谱-电喷雾电离质谱（LC-ESI-MS）从凝血酶激活的人血小板中鉴定磷酸化蛋白。
Electrophoresis. 1998 May;19(6):1015-23. doi: 10.1002/elps.1150190617.
引用本文的文献
1
Lysine 268 adjacent to transmembrane helix 5 of hamster P-glycoprotein is the major photobinding site of iodomycin in CHO B30 cells.仓鼠 P-糖蛋白跨膜螺旋 5 附近的赖氨酸 268 是碘霉素在 CHO B30 细胞中的主要光结合位点。
FEBS Open Bio. 2021 Apr;11(4):1084-1092. doi: 10.1002/2211-5463.13112. Epub 2021 Feb 28.
2
Large Scale Mass Spectrometry-based Identifications of Enzyme-mediated Protein Methylation Are Subject to High False Discovery Rates.基于大规模质谱法对酶介导的蛋白质甲基化进行鉴定时，假阳性率很高。
Mol Cell Proteomics. 2016 Mar;15(3):989-1006. doi: 10.1074/mcp.M115.055384. Epub 2015 Dec 23.
3
Structural cuticular proteins from arthropods: annotation, nomenclature, and sequence characteristics in the genomics era.节肢动物的结构表皮蛋白：基因组时代的注释、命名和序列特征。
Insect Biochem Mol Biol. 2010 Mar;40(3):189-204. doi: 10.1016/j.ibmb.2010.02.001. Epub 2010 Feb 18.
4
Identification of four novel types of in vitro protein modifications.四种新型体外蛋白质修饰的鉴定。
J Proteome Res. 2008 Oct;7(10):4603-8. doi: 10.1021/pr800456q. Epub 2008 Sep 4.
5
Cloning and expression of Tenebrio molitor antifreeze protein in Escherichia coli.黄粉虫抗冻蛋白在大肠杆菌中的克隆与表达
Mol Biol Rep. 2009 Mar;36(3):529-36. doi: 10.1007/s11033-008-9210-y. Epub 2008 Feb 7.
6
Analysis of electroblotted proteins by mass spectrometry: protein identification after Western blotting.通过质谱分析电印迹蛋白质：蛋白质印迹后的蛋白质鉴定
Mol Cell Proteomics. 2008 Feb;7(2):308-14. doi: 10.1074/mcp.M700415-MCP200. Epub 2007 Oct 15.
7
Use of nitrocellulose membranes for protein characterization by matrix-assisted laser desorption/ionization mass spectrometry.使用硝酸纤维素膜通过基质辅助激光解吸/电离质谱法进行蛋白质表征。
Anal Chem. 2006 Jul 15;78(14):5102-8. doi: 10.1021/ac060344t.
8
Extraction and characterization of adenovirus proteins from sodium dodecylsulfate polyacrylamide gel electrophoresis by matrix-assisted laser desorption/ionization mass spectrometry.
J Am Soc Mass Spectrom. 2000 Apr;11(4):356-61. doi: 10.1016/s1044-0305(00)00101-x.
9
Observation of gel-induced protein modifications in sodium dodecylsulfate [corrected] polyacrylamide gel electrophoresis and its implications for accurate molecular weight determination of gel-separated proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.十二烷基硫酸钠[校正后]聚丙烯酰胺凝胶电泳中凝胶诱导的蛋白质修饰观察及其对通过基质辅助激光解吸电离飞行时间质谱法准确测定凝胶分离蛋白质分子量的意义。
J Am Soc Mass Spectrom. 1999 Jun;10(6):512-20. doi: 10.1016/s1044-0305(99)00022-7.
10
Electrospray mass spectra from protein electroeluted from sodium dodecylsulfate polyacrylamide gel electrophoresis gels.从十二烷基硫酸钠聚丙烯酰胺凝胶电泳凝胶中电洗脱的蛋白质的电喷雾质谱。
J Am Soc Mass Spectrom. 1999 May;10(5):453-5. doi: 10.1016/S1044-0305(99)00012-4.
本文引用的文献
1
Mass spectrometric and Edman sequencing of lipocortin I isolated by two-dimensional SDS/PAGE of human melanoma lysates.
Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1927-31. doi: 10.1073/pnas.90.5.1927.
2
Electrospray ionization mass spectrometry on hydrophobic peptides electroeluted from sodium dodecyl sulfate-polyacrylamide gel electrophoresis application to the topology of the sarcoplasmic reticulum Ca2+ ATPase.十二烷基硫酸钠-聚丙烯酰胺凝胶电泳洗脱的疏水肽的电喷雾电离质谱法：在肌浆网Ca2+ATP酶拓扑学研究中的应用
Anal Biochem. 1993 Oct;214(1):50-7. doi: 10.1006/abio.1993.1455.
3
Combined plasma-desorption mass spectrometry and Edman degradation applied to simultaneous sequence determination of isoforms of structural proteins from the cuticle of Locusta migratoria.联合应用血浆解吸质谱法和埃德曼降解法同时测定飞蝗表皮结构蛋白的同工型序列。
Eur J Biochem. 1993 Oct 1;217(1):267-73. doi: 10.1111/j.1432-1033.1993.tb18242.x.
4
Analysis of recombinant Schistosoma mansoni antigen rSmp28 by on-line liquid chromatography-mass spectrometry combined with sodium dodecyl sulfate polyacrylamide gel electrophoresis.在线液相色谱-质谱联用结合十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析重组曼氏血吸虫抗原rSmp28
Anal Biochem. 1994 Jan;216(1):127-34. doi: 10.1006/abio.1994.1016.
5
Determination of the covalent structure of an N- and C-terminally blocked glycoprotein from endocuticle of Locusta migratoria. Combined use of plasma desorption mass spectrometry and Edman degradation to study post-translationally modified proteins.
Eur J Biochem. 1991 Jan 30;195(2):495-504. doi: 10.1111/j.1432-1033.1991.tb15730.x.
6
cDNA cloning and deduced amino acid sequence of a major, glycine-rich cuticular protein from the coleopteran Tenebrio molitor. Temporal and spatial distribution of the transcript during metamorphosis.
Eur J Biochem. 1992 Jun 15;206(3):813-9. doi: 10.1111/j.1432-1033.1992.tb16989.x.
7
Preincubation with cysteine prevents modification of sulfhydryl groups in proteins by unreacted acrylamide in a gel.用半胱氨酸预孵育可防止凝胶中未反应的丙烯酰胺对蛋白质中的巯基进行修饰。
Electrophoresis. 1992 Nov;13(11):882-4. doi: 10.1002/elps.11501301193.
8
Characterization of a cDNA clone encoding a glycine-rich cuticular protein of Tenebrio molitor: developmental expression and effect of a juvenile hormone analogue.
Insect Mol Biol. 1992;1(2):53-62. doi: 10.1111/j.1365-2583.1993.tb00105.x.
Zotero 插件