From: Subject: Protonic NanoMachine Group Date: Wed, 5 May 2004 08:25:07 -0700 MIME-Version: 1.0 Content-Type: multipart/related; boundary="----=_NextPart_000_0017_01C4327A.79258F00"; type="text/html" X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1106 This is a multi-part message in MIME format. ------=_NextPart_000_0017_01C4327A.79258F00 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Content-Location: http://www.fbs.osaka-u.ac.jp/en/seminar/09a.html Protonic NanoMachine Group


HOMEResearch=20 Divisions

3D"Laboratories

Professor NAMBA, Keiichi, = Ph.D. TEL +81-774-98-2543
Associate Prof. IMADA,=20 Katsumi, Ph.D. TEL +81-774-98-2543
Assistant Prof. YONEKURA, Koji, = Ph.D. TEL +81-774-98-2543
FAX +81-774-98-2575
Postal Mail Address Protonic NanoMachine = Project, ERATO,=20 JST 3-4 Hikaridai, Seika, Kyoto 619-0237=20 Japan

Protonic NanoMachine Group = aims at the=20 ultimate understanding of the mechanisms of = self-assembly and=20 its regulation, conformational switching, force = generation,=20 and energy transduction by biological macromolecular=20 complexes. By convergence of complementary techniques, = such as=20 X-ray diffraction and electron cryomicroscopy for=20 high-resolution analysis of three-dimensional = structures, and=20 optical and electronic measurements on individual = molecular=20 complexes for analyzing their dynamic behaviors, we = try to=20 reveal the basic principles behind their functional=20 mechanisms, in the hope that they will become a basis = for=20 artificial nanomachine design and nanotechnology.

1 Mechanism of = self-assembly and its=20 regulation

The bacterial flagellum, a motile organelle, = for=20 example, is a huge protein complex made of about 25 = proteins, each=20 of which forms either a ring or rod-like structure with a = range of=20 subunit copy numbers from a few to few tens of thousands. = The whole=20 structure spans from the cytoplasmic face of the cell = membrane to=20 the extra-cellular space, where the helical filament grows = up to=20 around 15 micrometer. The assembly proceeds one part after = another=20 from the base to the tip in an efficient and well-regulated = manner.=20 We try to reveal the regulatory mechanisms based on the = structure=20 and folding dynamics of individual component proteins and=20 sub-complexes.

2 Mechanism of = conformational=20 switching

Quick reversal of the flagellar motor rotation = and=20 polymorphic switching of the flagellar filament between = left- and=20 right-handed helical forms switch the direction of cell = swimming.=20 The universal joint function of the flagellar hook is = essential in=20 transmitting the torque to the long helical filament that = could be=20 oriented in various directions. All these processes involve = highly=20 precise and cooperative switching in the subunit protein=20 conformation coupled with switching in the interactions with = other=20 protein subunits. We try to visualize these conformational = switching=20 and understand the mechanisms of mechanical signal = transduction=20 based on the structure and = dynamics.

3 Torque generation=20 mechanism

The flagellar motors are only 30 to 40 nm in = diameter,=20 and yet, they rotate as fast as 20,000 to 100,000 rpm. We = try to=20 understand the mechanism of torque generation by studying = the=20 three-dimensional structure and dynamic behavior of its = rotation at=20 high spatial and temporal = resolution.
 
   

4 Energy transduction=20 mechanism
The high-speed rotation of the = flagellar=20 motor is powered by the flow of protons or Na ions through a = membrane channel protein complex that performs the stator = function=20 of the flagellar motor. The proton or ion flow is driven by = the=20 proton or ion motive force across the cytoplasmic membrane, = and the=20 current amounts only to several tens femtoampere. We try to = measure=20 this extremely small electric current precisely to obtain = clues to=20 the energy transduction mechanism.

5 Protein export=20 mechanism
The assembly of the flagellar = axial=20 structure with tens of thousands of protein subunits always = occurs=20 at the distal end of the growing flagellum. The component = proteins=20 are selectively exported from the cytoplasm into the narrow = central=20 channel of the flagellum by a protein complex attached on = the=20 cytoplasmic face of the motor. This flagellar protein export = apparatus uses the energy of ATP hydrolysis and is highly = homologous=20 to the type III protein secretion system of pathogenic = bacteria, by=20 which pathogenic effecter proteins are secreted into host = cells. We=20 try to visualize the export process by single molecule = technique and=20 understand the mechanism of selective export based on the=20 structure.

6 Design principles for = protein=20 nanomachine
Protein nanomachines have a = unique and=20 special ability to form three-dimensional structures and = large=20 complexes so that individual atoms take well-defined=20 three-dimensional positions to perform specific and yet = various=20 functions in a highly precise manner. This ability of=20 self-organization is a great advantage in nanotechnology=20 development, because, without this feature, mass production = of=20 nanomachines is impossible and therefore practical = applications=20 cannot be expected no matter how useful individually made=20 nanomachines could be. The outcome of our studies on protein = nanomachines, which work flexibly and precisely at the same = time, is=20 expected to produce much useful knowledge to eventually form = a basis=20 for design principles for artificial nanomachines of = practical=20 use.
=20
3D"Lab3DPublications=203D"Other

HOMEResearch=20 Divisions

(C) 2002=20 Osaka University Graduate School of Frontier=20 Biosciences

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