Citoplasma y citoesqueleto. Motilidad celular.

Estructura de cilios y flagelos

ESME110

Metabolismo oxidativo

Organizacin ms compleja:

2.7 billones

- organelas membranosas - citoesqueleto Eficiencia energtica

3.8 billones

Evolution of cells Present-day cells evolved from a common prokaryotic ancestor along three lines of descent, giving rise to archaebacteria, eubacteria, and eukaryotes. Mitochondria and chloroplasts originated from the endosymbiotic association of aerobic bacteria and cyanobacteria, respectively, with the ancestors of eukaryotes.

The major features of eucaryotic cells. The drawing depicts a typical animal cell, but almost all the same components are found in plants and fungi and in single-celled eucaryotes such as yeasts and protozoa. Plant cells contain chloroplasts in addition to the components shown here, and their plasma membrane is surrounded by a tough external wall formed of cellulose.

Citosol Agua: 75 a 80% Electrolitos: K, Mg, P, S, Na, Cl, Ca,etc Protenas: 10 a 15% Lpidos Carbohidratos: glucgeno

Hypotheses for the evolutionary origins of some membrane-bounded organelles. The origins of mitochondria, chloroplasts, ER, and the cell nucleus could explain the topological relationships of these intracellular compartments in eucaryotic cells. (A) A possible pathway for the evolution of the cell nucleus and the ER. In some bacteria the single DNA molecule is attached to an invagination of the plasma membrane, called a mesosome. Such an invagination in a very ancient procaryotic cell could have spread to form an envelope around the DNA while still allowing access of the DNA to the cell cytosol (as is required for DNA to direct protein synthesis). This envelope is presumed to have eventually pinched off completely from the plasma membrane, producing a nuclear compartment surrounded by a double membrane. As illustrated, the nuclear envelope is organized by a fibrous shell called the nuclear lamina and is penetrated by communicating channels called nuclear pore complexes. Because it is surrounded by two membranes that are in continuity where they are penetrated by these pores, the nuclear compartment is topologically equivalent to the cytosol. The lumen of the ER is continuous with the space between the inner and outer nuclear membranes and topologically equivalent to the extracellular space. (B) Mitochondria (and chloroplasts) are thought to have originated when a bacterium was engulfed by a larger pre-eucaryotic cell. They retain their autonomy. This may explain why the lumens of these organelles remain isolated from the vesicular traffic that interconnects the lumens of many other intracellular compartments.

 200 tipos celulares diferentes 5 tipos principales de tejidos: (1) epitelial; (2) conectivo; (3) muscular; (4) nervioso; (5) sangre 5 compartimentos intracelulares: (1) el ncleo y el citosol; (2) organelas que funcionan en la va secretoria y endoctica; (3) mitocondria; (4) cloroplastos (c. vegetal); (5) peroxisomas Las clulas NO pueden sintetizar organelas de novo. Cada clula tiene un desarrollo de organelas adecuado a su funcin.

Todas las protenas se sintetizan en el citosol Ribosomas asociados al RE Ribosomas libres (polisomas)

i) Protenas que quedan asociadas a las membranas (protenas integrales, ancladas y asociadas: receptores, canales, etc.). ii) Protenas que quedan en el lumen del RER o el Golgi. iii) Protenas destinadas a otras organelas (lisomas). iv) Protenas secretorias.

El destino de la proteina esta determinado por su secuencia:

citoesqueleto
Red de filamentos proteicos, que confieren forma y organizacin interna a la clula y permiten su movimiento (total y el de organelas). Estructura dinmica 1.- Microfilamentos (actina) 2.- Filamentos intermedios 3.- Microtbulos (tubulina) Protenas accesorias

Polmeros de actina 7 nm de dimetro Concentrados en la periferia de la clula Se organizan en estructuras complejas (redes tridimensionales) Contribuyen al mantenimiento de la forma y polaridad celular Forman pseudopodios y microvellosidades (migrar y fagocitar compuestos) Forman uniones clulaclula y clula-matriz extracelular (sealizacin intracelular)

Polimerizacin reversible, ATP

ATP ATP ATP ATP

ATP

ADP ADP ATP

43 kDa

Frmacos: Citocalasina.- se une al extremo +end y bloquea la elongacin

Faloidina.- se une a los filamentos de actina e inhibe su disociacin

Filamento polar

Protenas de unin a actina Profilina .- favoroce la polimerizacin Cofilina.- favorece el desemsamblaje Arp2/3 (sitios de nucleacin=inicio del ensamblaje) Distrofina (distrofia muscular)

Assembly and structure of actin filaments (A) Actin monomers (G actin) polymerize to form actin filaments (F actin). The first step is the formation of dimers and trimers, which then grow by the addition of monomers to both ends. (C, courtesy of Dan Richardson.) Reversible polymerization of actin monomers

Los filamentos de actina se organizan en redes

Actin networks and filamin Filamin is a dimer of two large (280-kd) subunits, forming a flexible Vshaped molecule that crosslinks actin filaments into orthogonal networks. The carboxy-terminal dimerization domain is separated from the aminoterminal actin-binding domain by repeated -sheet spacer domains.

Organization of microvilli The core actin filaments of microvilli are crosslinked into closely packed bundles by fimbrin and villin. They are attached to the plasma membrane along their length by lateral arms, consisting of myosin I and calmodulin. The plus ends of the actin filaments are embedded in a cap of unidentified proteins at the tip of the microvillus.

Cell surface projections involved in phagocytosis and movement. Lamelopodios, pseudopodos, filopodios

Attachment of stress fibers to the plasma membrane at focal adhesions Focal adhesions are mediated by the binding of integrins to proteins of the extracellular matrix. Stress fibers (bundles of actin filaments crosslinked by -actinin) are then bound to the cytoplasmic domain of integrins by complex associations involving a number of proteins. Two possible associations are illustrated: 1) talin binds to both integrin and vinculin, which in turn binds to actin, and 2) integrin binds to actinin. A number of other proteins (not shown) are also present at focal adhesions and may be involved in anchoring stress fibers to the plasma membrane.

 Polmeros de protenas fibrosas (50) 8-11 nm de dimetro Contribuyen a la estabilidad mecnica de las clulas animales Forman uniones clulaclula y clula-matriz extracelular Tipo I, II: queratinas Tipo III: vimentina Tipo IV: neurofilamentos Tipo V: lamininas nucleares

A pesar de la diversidad, las protenas que forman FI tienen una estructura comn conservada

Structure of intermediate filament proteins Intermediate filament proteins contain a central -helical rod domain of approximately 310 amino acids (350 amino acids in the nuclear lamins). The N-terminal head and C-terminal tail domains vary in size and shape.

Assembly of intermediate filaments The central rod domains of two polypeptides wind around each other in a coiled-coil structure to form dimers. Dimers then associate in a staggered antiparallel fashion to form tetramers. Tetramers associate end to end to form protofilaments and laterally to form filaments. Each filament contains approximately eight protofilaments wound around each other in a ropelike structure. Phosphorylation.

Experimental demonstration of keratin function A plasmid encoding a mutant keratin that interferes with the normal assembly of keratin filaments was microinjected into one pronucleus of a fertilized egg. Microinjected embryos were then transferred to a foster mother, and some of the offspring were found to have incorporated the mutant keratin gene into their genome. Expression of the mutant gene in these transgenic mice disrupted the keratin cytoskeleton of cells of the epidermis, resulting in severe skin blistering due to cell lysis following mild mechanical stress

Esclerosis lateral amiotrfica (ALS). Enfermedad de Lou Gehring (Stephen Hawking). Acumulacin y ensamblaje anormal de neurofilamentos.

Uniones celulares

Attachment of intermediate filaments to desmosomes and hemidesmosomes (A) Electron micrograph illustrating keratin filaments (arrows) attached to the dense plaques of intracellular protein on both sides of a desmosome. (B) Schematic of a desmosome. Intermediate filaments are anchored to sites of cell-cell adhesion by desmoplaskin. (C) Schematic of a hemidesmosome. Intermediate filaments are anchored to an integrin by plectin. (A, Don Fawcett/Photo Researchers, Inc.

 Componentes ms grandes del citoesqueleto Cilindros huecos de aprox. 25 nm de dimetro polmeros de y tubulina (isoformas)
Participan: -Transporte intracelular - Cilios y flagelos - Divisin celular

GDP GDP GTP GTP 37C Mg2+ GTP

_
Structure of microtubules Dimers of - and -tubulin polymerize to form microtubules, which are composed of 13 protofilaments assembled around a hollow core. Polimerizacin reversible MT= estructuras polares. Polo - y polo +. Cada monmero de tubulina une dos molculas de GTP, e hidroliza una y la energa liberada se utiliza para el autoensamblaje de otros monmeros La nucleacin de un MT es lenta; su elongacin es rpida. La nucleacin sucede en una regin especfica de la clula=centrosoma=centro organizador de MT

MT MT

MT

MT

centrosoma
Centriolos (L) Material pericentriolar

MT

Growth and shrinkage in a microtubule array. The array of microtubules anchored in a centrosome is continually changing, as new microtubules grow ( red arrows) and old microtubules shrink ( blue arrows).

MT

+ +

/ tubulina tubulina Protenas ..

Dynamic instability of microtubules Dynamic instability results from the hydrolysis of GTP bound to -tubulin during or shortly after polymerization, which reduces its binding affinity for adjacent molecules. Growth of microtubules continues as long as there is a high concentration of tubulin bound to GTP. New GTP-bound tubulin molecules are then added more rapidly than GTP is hydrolyzed, so a GTP cap is retained at the growing end. However, if GTP is hydrolyzed more rapidly than new subunits are then added, the presence of GDP-bound tubulin at the end of the microtubule leads to disassembly and shrinkage. Only the plus ends of microtubules are illustrated.

Postranslation modifications.- Acetylation. Detyrosination (MT maduration): (1) clock; (2) sitios de unin a MAPs

Protenas asociadas a microtbulos (MAPs)

Se unen a las subunidades de tubulina de los MT - Estabilizan y desestabilizan MT - Guian a los MT a regiones celulares especficas - Median las interacciones de los MT con otras protenas en las clulas

Tipo I: MAP1 Tipo II: MAP2, MAP4 and tau (Alzheimer) Motores: miosina, dineina,kinesina

axn

MT

MT y MAPs ayudan a crear citoplasma diferenciado funcionalmente

p.ej neurona

Organization of microtubules in nerve cells Two distinct types of processes extend from the cell body of nerve cells (neurons). Dendrites are short processes that receive stimuli from other nerve cells. The single long axon then carries impulses from the cell body to other cells, which may be either other neurons or an effector cell, such as a muscle. Stable microtubules in both axons and dendrites terminate in the cytoplasm rather than being anchored in the centrosome. In dendrites, microtubules are oriented in both directions, with their plus ends pointing both toward and away from the cell body. In contrast, all of the axon microtubules are oriented with their plus ends pointing toward the tip of the axon.

Microtubule motor proteins Kinesin and dynein move in opposite directions along microtubules, toward the plus and minus ends, respectively. Kinesin consists of two heavy chains, wound around each other in a coiled-coil structure, and two light chains. The globular head domains of the heavy chains bind microtubules and are the motor domains of the molecule. Dynein consists of two or three heavy chains (two are shown here) in association with multiple light and intermediate chains. The globular head domains of the heavy chains are the motor domains.

Transporte axonal antergrado (Kinesina) - Rpido. 400 mm/dia. Transporte de organelas en forma saltatoria (mitocodrias, vesiculas y granulos). -lento. El axoplasma (citoesqueleto y proteinas solubles) se transporta lentamente. Dos componentes cinticos:

a) el lento se mueve a un ritmo de de 0.22.5 mm/dia, y transporta las formas solubles de las proteinas del citoesqueleto. b) el ms rapido se mueve a 5 mm/dia, y transporta una mezcla de proteinas: actina, clatrina, enzimas, etc.
Transporte retrgrado (dineina)

Es rpido (aprox. 200 mm/dia). Transporta materiales desde la terminal hasta el soma para degradacin o reutilizacin. Son paquetes rodeados de membranas formados por endocitosis y que pertenecen al sistema lisosomal.

Intracellular organization of microtubules The minus ends of microtubules are anchored in the centrosome. In interphase cells, the centrosome is located near the nucleus and microtubules extend outward to the cell periphery. During mitosis, duplicated centrosomes separate and microtubules reorganize to form the mitotic spindle. Colchicina.- previene la polimerizacin. Taxol.- se une a los MT y los estabiliza; impide la despolimerizacin.

Cilios (0.25 m) y flagelos (200 m)

Cilios 9+2, mviles

A) Computer-enhanced electron micrograph of a cross section of the axoneme of a rat sperm flagellum; (c) basal body

9+0, sensores

Structure of the axoneme of cilia and flagella. (B) Schematic cross section of an axoneme. The nine outer doublets consist of one complete (A) and one incomplete (B) microtubule, containing only 10 or 11 protofilaments. The outer doublets are joined to each other by nexin links and to the central pair of microtubules by radial spokes. Each outer microtubule doublet is associated with inner and outer dynein arms. (A, K. G. Murti/Visuals Unlimited.)

El movimiento de los cilios y flagelos resulta del deslizamiento de los MT, consecuencia de la hidrlisis de ATP por la dineina

1.- When dynine arms use ATP, microtubules doublets slide past each other

2.- When microtubules slide past each other the flagella changes shape

3.- When flagella or cilia change shape the cell moves

El mal funcionamiento de cilios y flagelos produce enfermedades

Las clulas eucariotas muestran diferentes niveles de organizacin: (1) compartimentos membranosos; (2) citoesqueleto

Proteins of the cytoskeleton Microfilaments Actins - Actin-binding proteins - Actinin - Arp2/3 complex Cofilin - Destrin - Gelsolin - Myosins - Profilin - Tropomodulin Troponin (T, C, I) - Tropomyosin - Wiskott-Aldrich syndrome protein type 1 and 2 (Cytokeratin, type I, type II) - type 3 (Desmin, GFAP, Peripherin, Vimentin) - type 4 (Internexin, Nestin, Neurofilament) - type 5 (Lamin A, B) Dyneins - Kinesins - MAPs (Tau protein, Dynamin) - Tubulins Alpha catenin - Beta catenin - Plakoglobin (gamma catenin) Delta catenin APC - Dystrophin (Dystroglycan) - plakin (Desmoplakin, Plectin) - Spectrin - Talin - Utrophin - Vinculin

Intermediate filaments Microtubules Catenins

Other