Main description:

This volume aims at providing an update on state of the art methodologies to study various aspects of selenoprotein biology. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Selenoproteins: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

Part I: Bioinformatic Tools for Selenoprotein Identification and Evolution

1. SECISearch3 and Seblastian: In Silico Tools to Predict SECIS Elements and Selenoproteins

Marco Mariotti

2. Selenoprofiles: A Computational Pipeline for Annotation of Selenoproteins

Didac Santesmasses, Marco Mariotti, and Roderic Guigo

3. SelGenAmic: An Algorithm for Selenoprotein Gene Assembly

Liang Jiang and Qiong Liu

Part II: Selenoprotein Biosynthesis

4. Selenocysteine tRNA[Ser]Sec, the Central Component of Selenoprotein Biosynthesis: Isolation, Identification, Modification and Sequencing

Bradley A. Carlson, Byeong Jae Lee, Petra A. Tsuji, Paul R. Copeland, Ulrich Schweizer, Vadim N. Gladyshe, and Dolph L. Hatfield

5. Identification and Characterization of Proteins that Bind to Selenoprotein 3' UTRs

Eric M. Cockman and Donna M. Driscoll

6. Specific Chemical Approaches for Studying Mammalian Ribosomes Complexed with Ligands Involved in Selenoprotein Synthesis

Olga Kossinova, Alexey Malygin, Alain Krol, and Galina Karpova

7. In vitro Translation Assays for Selenocysteine Insertion

Mark H. Pinkerton and Paul R. Copeland

8. Studying Selenoprotein mRNA Translation using RNA-Seq and Ribosome Profiling

Brian K. Dalley, Lisa Baird, and Michael T. Howard

9. Modification of Selenoprotein mRNAs by Cap Tri-methylation

Anne-Sophie Gribling-Burrer, Gilbert Eriani, and Christine Allmang

Part III: Selenoprotein Detection and Quantification

10. Total Selenium Quantification in Biological Samples by Inductively Coupled Plasma Mass Spectrometry (ICP-MS)

Veronique Vacchina and Jean Dumont

11. Quantification of SeMet and SeCys in Biological Fluids and Tissues by Liquid Chromatography Coupled to Inductively Coupled Plasma Mass Spectrometry (HPLC - ICP MS)

Veronique Vacchina, Katarzyna Bierla, Joanna Szpunar, and Ryszard Lobinski

12. Simultaneous Speciation of Selenoproteins and Selenometabolites in Plasma and Serum

Belen Callejon-Leblic, Gema Rodriguez-Moro, Tamara Garcia-Barrera, and Jose Luis Gomez-Ariza

13. Radioactive 75Se Labeling and Detection of Selenoproteins

Sun Hee Yim, Ryuta Tobe, Anton A. Turanov, and Bradley A. Carlson

14. Non-radioactive Isotopic Labeling and Tracing of Selenoproteins in Cultured Cell Lines

Jordan Sonet, Sandra Mounicou, and Laurent Chavatte

15. Detection of Selenoproteins by Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP MS) in Immobilized pH Gradient (IPG) Strips

Jordan Sonet, Sandra Mounicou, and Laurent Chavatte

16. Imaging of Selenium by Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) in 2-D Electrophoresis Gels and Biological Tissues

Elisa Castaneda Santa Cruz, J. Susanne Becker, J. Sabine Becker, and Alessandra Sussulini

Part IV: Selenoprotein Overexpression, Purification, and Activity

17. Overexpression of Recombinant Selenoproteins in E. coli

Qing Cheng and Elias S.J. Arner

18. Preparation of Selenocysteine-containing Forms of Human SELENOK and SELENOS

Zhengqi Zhang, Jun Liu, and Sharon Rozovsky

19. Selenocysteine Mediated Expressed Protein Ligation of SELENOM

Jun Liu, Qingqing Chen and Sharon Rozovsky

20. Monitoring of Methionine Sulfoxide Content and Methionine Sulfoxide Reductase Activity

Lionel Tarrago, Emmanuel Oheix, Zalan Peterfi, and Vadim N. Gladyshev

21. Selective Evaluation of Thioredoxin Reductase Enzymatic Activities

Elias S.J. Arner

Part V: In Vivo Studies

22. Association of Single Nucleotide Polymorphisms in Selenoprotein Genes with Cancer Risk

Catherine Meplan

23. Identification of Genetic Disorders Causing Disruption of Selenoprotein Biosynthesis

Erik Schoenmakers and Krishna Chatterjee