This book describes a new family of bio-polymergels made from cytoskeletal proteins - actin, microtubule, andtropomyosin. The importance of the gelstate with multi-scale hierarchical structure is emphasized to utilize emergentfunctions in living organisms. Detailed protocol of gel preparation, specifiedmethod of structure investigation, and dynamic studies of self-organization,self-healing, synchronized oscillating, and autonomous motility functions areintroduced together with biomimeticfunctions of synthetic hydrogels.
1 Introduction1-1 What is Hydrogel? Natural and Synthetic Polymer Hydrogels Basic Structure and Behaviors of Polymer Hydrogels1-2 Self-Assembly and Dynamic Structure of Cytoskeletal Filaments1-3 Why Cytoskeletal Gel? --- Super -Giant and Multi-Scale Hierarchical Structure 2 Microtubules Gel2-1 Preparation of Microtubules Gel2-2 Supra-Molecular Structure of Microtubules Gel2-3 Specific Assembly Process of Chemically Modified Tublin2-4 Photo-Chemical Catalytic Activity of Microtubule Gel2-5 Emergent Functions as Protein Gel Motors3 Actin Gel3-1 Preparation of Actin Gel3-2 Supra-Molecular Structure and Emergent Functions of Actin Gel4 Tropomyosin Gel4-1 Preparation and properties of tropomyosin Gel4-2 Superior Cell Penetration of Synthetic Tropomyosin
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1 | (6) |
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1 | (1) |
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1.2 Chemical Gel, Physical Gel, and Biological Gel |
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2 | (2) |
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1.3 Living Organism Consists of Hydrogels |
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4 | (3) |
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5 | (2) |
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7 | (14) |
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2.1 Self-Assembly and Dynamic Structure of Cytoskeletal Filaments |
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7 | (5) |
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8 | (1) |
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8 | (3) |
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11 | (1) |
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2.2 Multi-Scale Hierarchy: Origin of Emergence |
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12 | (1) |
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2.3 Cytoskeletal Protein Gels: Multi-Scale Hierarchical Supra-Macromolecular Gel (MHSMG) |
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13 | (3) |
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13 | (2) |
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15 | (1) |
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2.4 Bio-Motor Gel with Emergent Function |
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16 | (5) |
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18 | (3) |
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21 | (14) |
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3.1 Preparation of Actin Gel |
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21 | (4) |
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3.1.1 Preparation of Acetone Powder of Striated Muscle |
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21 | (1) |
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3.1.2 Extraction of Actin from Acetone Powder |
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22 | (1) |
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23 | (2) |
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3.2 Emergent Functions of Actin Gel |
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25 | (10) |
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3.2.1 Mechanical Property |
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25 | (2) |
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3.2.2 Reversible Sol--Gel Transition |
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27 | (1) |
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3.2.3 Self-Healing Function |
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27 | (4) |
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3.2.4 Autonomous Oscillation |
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31 | (2) |
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33 | (2) |
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35 | (24) |
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4.1 Preparation of Microtubule Gel |
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35 | (5) |
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4.1.1 Tubulin Purification |
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35 | (2) |
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4.1.2 MT Gel Formation by Cross-Linking |
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37 | (3) |
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4.2 Supramolecular Structure of Microtubule Gel |
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40 | (5) |
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4.2.1 Direct Observation of MT Network |
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40 | (1) |
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4.2.2 Mechanical Property |
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41 | (2) |
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4.2.3 Reversible Sol--Gel Transition |
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43 | (2) |
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4.3 Emergent Functions as Protein Gel Motors |
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45 | (9) |
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4.3.1 Motility Assay of MT Gel |
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45 | (4) |
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4.3.2 Enhanced Motion of MT Gel |
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49 | (4) |
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4.3.3 Role of Cross-Linkage on MT Gel Motility |
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53 | (1) |
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4.4 Specific Assembly Processes of Chemically Modified Tubulin |
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54 | (5) |
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4.4.1 Photo-Enhanced Polymerization of Ru(bpy)32+-Tubulin Conjugate |
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54 | (1) |
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4.4.2 Teardrop Pattern Formation of MTs in a Hydrodynamic Flow |
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55 | (1) |
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56 | (3) |
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59 | (12) |
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5.1 Preparation of Tropomyosins |
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59 | (4) |
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5.1.1 Preparation of Tropomyosin from Striated Muscle |
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59 | (1) |
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5.1.2 Preparation of Recombinant Tropomyosin |
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60 | (1) |
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5.1.3 Gelation of Tropomyosin |
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61 | (1) |
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5.1.4 Cell-Penetrating Assay of Tropomyosin Derivatives |
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62 | (1) |
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5.2 pH Responsive Swelling and Contracting Behaviors of Tropomyosin Gel |
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63 | (1) |
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5.2.1 pH Response of Tropomyosin Gel |
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63 | (1) |
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5.3 Superior Cell Penetration of Tropomyosin Derivatives |
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64 | (7) |
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5.3.1 Design of Coiled-Coil Cell-Penetrating Proteins |
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64 | (1) |
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5.3.2 Cell-Penetrating Activity of Tropomyosin Derivatives |
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65 | (1) |
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5.3.3 Effect of Molecular Length on Cell Penetration |
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66 | (1) |
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5.3.4 Cell-Penetrating Activity of Non-cationic Tropomyosin Derivatives |
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67 | (2) |
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69 | (2) |
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71 | (2) |
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7 Biomimetic Functions of Synthetic Polymer Gels |
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73 | |
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7.1 Soft and Wet Artificial Muscles |
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73 | (2) |
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75 | (1) |
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75 | (1) |
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7.4 Strong Hydrogels with Extremely Low Friction |
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75 | (1) |
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7.5 Electroconductive Hydrogel |
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76 | (1) |
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7.6 Tunable Photonic Crystal |
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Professor Yoshihito Osada obtained his Bachelors degree in chemistry from Waseda University, Japan, and received his Ph.D. in polymer science from Moscow State University (supervisor: Prof. V.A. Kabanov). He began as a professor in 1992, then became the Dean and eventually the Vice President of Hokkaido University, Sapporo, Japan. He was invited to be Deputy Director at the Advanced Science Institute, RIKEN in 2007. He is currently a Senior Visiting Scientist at RIKEN and a professor emeritus at Hokkaido University.
Professor Yoshihito Osada is a pioneer of polymer gels. He developed artificial muscle systems using various functional gels such as Shape Memory Gel and Double Network Gel with excellent mechanical performances. His current interests focus on highly-hierarchical protein gels with emergent muscle functions, nano-patterning of the gel, and electro-conductive gels.
Ken-Ichi Sano received his B.S. degree in biologyfrom Osaka City University, Japan, in 1993, and his Ph.D. degree in biophysics from Nagoya University, Japan, in 2000. In 1994, he joined the International Institute for Advanced Research, Panasonic, and since 1999 he has worked for RIKEN, Harima Institute at SPring-8. Since 2003 he has been with the Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, where he studied interfacial molecules between biomaterials and inorganics. In 2008, he joined the Molecular and System Life Science Unit, Advanced Science Institute, RIKEN as a Contract Researcher, and became Deputy Unit Leader in 2009 where he studied hydrogels of cytoskeletal proteins. In April 2011 he joined the Department of Innovative Systems Engineering, Nippon Institute of Technology as an Associate Professor. His current research interests include cellular drug delivery systems and evaluation of antidepressant agents using novel models.
Ryuzo Kawamura wasborn in Nara, Japan, in 1980. He obtained his Bachelors degree in bioengineering from Tokyo Institute of Technology, Japan, in 2005 and received his Ph.D. in polymer science from Hokkaido University, Japan (supervisor: Prof. J.P. Gong) in 2008. He did postdoctoral research at RIKEN and at the National Institute of Advanced Science and Technology (AIST). Currently, he is an Assistant Professor in Department of Chemistry, Saitama University, Japan.
In 2009, he joined the Molecular and System Life Science Unit, Advanced Science Institute, RIKEN. There he developed the Multi-scale Hierarchical Supra-Macromolecular Gels (MHSMG) as a new type of material. His current research interest is in coordinative and mesoscale functions of the cytoskeletal and motor proteins with both viewpoints of material science and biology.
