Mucins: Methods and Protocols

ISBN-13: 9781071636695 / Angielski

Akihiko Kameyama

Spis treści

Preface…
Table of Contents…
Contributing Authors…

Part I Extraction and Separation

1. Preparation of Jellyfish Mucins
Kiminori Ushida, Hiroshi Inui, Takuma Kaneko, Shinra Tanaka, Anri Mochizuki, Shiori Kaise, and Minami Sugiyama

2. Extraction of Mucin from Rodent Feces and Determination of O-Linked Oligosaccharide Chain Equivalent Derived from Fecal Mucin
Naomichi Nishimura, Hiroki Tanabe, and Tatsuya Morita

3. Preparation of Soluble Mucin Solutions from the Salivary Glands
Takanori Sugiura and Akihiko Kameyama

4. Isolation of Membrane Bound Mucins from Human Bronchial Epithelial Cells
Jerome Carpenter and Mehmet Kesimer

5. Extraction and Fractionation of Human Gastric Mucins from Gastric Juice
Rei Kawashima, Daigo Tsubokawa, Katsunori Iijima, and Takafumi Ichikawa

6. Extraction of Mucins from the Mammalian Intestinal Tract
Shota Okamoto, Mugen Taniguchi, and Ryu Okumura

7. Supported Molecular Matrix Electrophoresis
Akihiko Kameyama

Part II Staining, Detection, and Quantitation

8. Supported Immunohistochemical Staining of Mucins
Michiyo Higashi

9. Succinylation-Alcian Blue Staining of Mucins on Polyvinylidene Difluoride Membrane
Weijie Dong and Akihiko Kameyama

10. Quantitation of Mucin by Densitometry of an Alcian Blue-Stained Membrane
Mayumi Tamura and Yoichiro Arata

11. Quantitation of MUC5AC and MUC5B by Stable Isotope Labeling Mass Spectrometry
Giorgia Radicioni and Mehmet Kesimer

Part III Preparation and Analysis of Mucin Glycans

12. Preparation of O-Glycans from Mucins using Hydrazine Treatment
Yukinobu Goso and Makoto Kurihara

13. Eliminative Oximation of O-glycans from Mucins
Akihiko Kameyama

14. 9-Fluorenylmethyl Chloroformate Labeling for O-Glycan Analysis
Keita Yamada

15. Liquid Chromatography and Capillary Electrophoresis Analysis of 2AA-Labeled O-Glycans
Keita Yamada

16. Preparation of Mucin Glycopeptides by Organic Synthesis
Izuru Nagashima and Hiroki Shimizu

17. MALDI-TOF MS/MS Analysis of Permethylated O-Glycan Alditols Released from Mucins
Yukinobu Goso and Makoto Kurihara

18. Structural Elucidation of Sialylated O-Glycan Alditols Obtained from Mucins by Mass Spectrometry
Daigo Tsubokawa, Rei Kawashima, and Takafumi Ichikawa

19. Differential Glycoform Analysis of MUC1 Derived from Biological Specimens using an Antibody-Overlay Lectin Microarray
Atsushi Matsuda, Patcharaporn Boottanun, Sachiko Koizumi, Misugi Nagai, and Atsushi Kuno

20. ISOGlyP: O-Glycosylation Site Prediction using Peptide Sequences and GALNTs
Luisa Gracia Mazuca and Jonathon E. Mohl

Part IV Molecular Biology

21. Assessment of Mucin-Associated Gene Expression Levels on the Ocular Surface
Jun Shoji and Satoru Yamagami

22. Methylation Specific Electrophoresis
Seiya Yokoyama, Kei Matsuo, and Akihide Tanimoto

23. Expression Analysis of Genes Corresponding to Mucins and their Glycans from Cervical Tissue using RNA Sequencing
Sean Fair and Laura Abril-Parreño

24. Recombinant Production of Glycoengineered Mucins in HEK293-F Cells
Ling-Ting Huang, Marshall J. Colville, and Matthew Paszek

Part V Interaction of Mucins and Other Biomolecules

25. Analysis of the Interaction between Mucin and Green Fluorescent Protein (GFP)-Tagged Galectin-2 using a 96-Well Plate
Mayumi Tamura and Yoichiro Arata

26. Solution NMR Analysis of O-Glycopeptide-Antibody Interaction
Ryoka Kokubu, Shiho Ohno, Noriyoshi Manabe, and Yoshiki Yamaguchi

Part VI Mucin and Microorganism

27. Cultivation of Microorganisms in Media Supplemented with Mucin Glycoproteins
Hiromi Takada, Takane Katayama, and Toshihiko Katoh

28. Bacterial Enzyme Assay for Mucin Glycan Degradation
Toshihiko Katoh and Hisashi Ashida

29. Measurement of Mucinase Activity
Hiroki Tanabe, Tatsuya Morita, and Naomichi Nishimura

30. Adhesion Inhibition Assay for Helicobacter pylori to Mucin by lactobacillus
Keita Nishiyama and Takao Mukai

Part VII Imaging and MD Simulation of Mucins

31. Imaging of Mucin Networks with Atomic Force Microscopy
Jerome Carpenter and Mehmet Kesimer

32. Molecular Dynamics Simulation and Docking of MUC1 O-Glycopeptide
Ryoka Kokubu, Shiho Ohno, Noriyoshi Manabe, and Yoshiki Yamaguchi

Part VIII Mucin-Hydrogel and Physicochemical Properties

33. Fabrication and Characterization of Mucin Nanoparticles for Drug Delivery Applications
Ceren Kimna, Theresa M. Lutz, and Oliver Lieleg

34. Evaluation of Rheological Properties of Saliva by Determining the Spinnbarkeit
Taro Mukaibo and Mikio Yamada

35. Mechanical Characterization of Mucus on Intestinal Tissues by Atomic Force Microscopy
Momoka Horikiri, Mugen Taniguchi, Hiroshi Y. Yoshikawa, Ryu Okumura, and Takahisa Matsuzaki

Subject Index List…

Blurb

This volume explores the latest advancements in mucin research. The chapters in this book are organized into 8 parts and cover a wide range of topics such as mucin extraction, isolation, physicochemical property analysis, and experimental methods. The chapters also discuss the origins of mucins in jellyfish, feces, saliva and salivary glands, bronchi, stomach, and cervical tract; organic synthesis of peptides glycosylated at specific sites; analysis of mucin gene expression and methylation-specific electrophoresis of genes; imaging of mucin networks by AFM; and experimental methods using supported molecular matrix electrophoresis. 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.

Cutting-edge and comprehensive, Mucins: Methods and Protocols is a valuable resource for researchers who want to obtain in-depth insight on how to more thoroughly conduct mucin research in their laboratories.