title: Chapter 3 - Construction of cell-sized liposomes encapsulating actin and actin-cross-linking proteins.
abstract: To shed light on the mechanism underlying the active morphogenesis of living cells in relation to the organization of internal cytoskeletal networks, the development of new methodologies to construct artificial cell models is crucial. Here, we describe the successful construction of cell-sized liposomes entrapping cytoskeletal proteins. We discuss experimental protocols to prepare giant liposomes encapsulating desired amounts of actin and cross-linking proteins including molecular motor proteins, such as fascin, alpha-actinin, filamin, myosin-I isolated from brush border (BBMI), and heavy meromyosin (HMM). Subfragment 1 (S-1) is also studied in comparison to HMM, where S-1 and HMM are single-headed and double-headed derivatives of conventional myosin (myosin-II), respectively. In the absence of cross-linking proteins, actin filaments (F-actin) are distributed homogeneously without any order within the liposomes. In contrast, when actin is encapsulated together with an actin-cross-linking protein, mesh structures emerge that are similar to those in living motile cells. Optical microscopic observations on the active morphological changes of the liposomes are reported.
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Japan.
authors.forename: Kingo
authors.initials: K
authors.lastname: Takiguchi
authors.forename: Ayako
authors.initials: A
authors.lastname: Yamada
authors.forename: Makiko
authors.initials: M
authors.lastname: Negishi
authors.forename: Makoto
authors.initials: M
authors.lastname: Honda
authors.forename: Yohko
authors.initials: Y
authors.lastname: Tanaka-Takiguchi
authors.forename: Kenichi
authors.initials: K
authors.lastname: Yoshikawa
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D048429:Cell Size; D007364:Intercalating Agents / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000138:chemical synthesis / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D011485:Protein Binding
chemical_list: D000199:Actins; D007364:Intercalating Agents; D008081:Liposomes