研细胞信息转导.ppt

,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,Company Logo,*,单击此处编辑母版标题样式,LOGO,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,Company Logo,*,单击此处编辑母版标题样式,LOGO,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,Company Logo,*,单击此处编辑母版标题样式,LOGO,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,Company Logo,*,单击此处编辑母版标题样式,LOGO,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,Company Logo,*,单击此处编辑母版标题样式,LOGO,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,单击此处编辑母版标题样式,每天您发出多少,EMAILS？,每天您又收到多少手机短信呢？,History of signaling trnaduction pathway,In,1955,，,Sutherland,，,cAMP,second messenger，won Nobel prize in,1971,.,In,1963,，,cGMP,intracelluar messenger theory,In,1978,，,Rasmussen,，,Ca,2+,acting as,second messenger,In,1983,，,IP,3,and,DG,serving as,second messenger,In,80,s，,Gilman,and,Rodbell,，,G,protein,，won Nobel prize in,1994,80-90,，,tyrosine kinase and,signaling tranduction pathway,As early as 1938,Carl Cori,and,Getty Cori,found,phosphorylase existed in two forms:,the less active phosphorylase,b,;,and,the more active phosphorylase,a,.,In 1956,Edwin Krebs,and,Edmond Fischer,reported that a,“,converting enzyme,”could convert phosphorylase,b,to,phosphorylase,a.,Three years later,Krebs and Fischer,demonstrated that the conversion of phosphorylase,b,to,phosphorylase a,involved,covalent phosphorylation.,Krebs EG,Fischer E,.,The phosphorylase b to a converting enzyme of rabbit,skeletal muscle,.,Biochim Biophys Acta,1956;20:150-7.,The model proposed by Sutherland,In 1968,E Krebs lab found cAMP-dependent protein,Kinase(PKA).,Walsh DA,Perkins JP,and,Krebs EG,.,An Adenosine 3,5-Monophosphate-dependant Protein Kinase from Rabbit Skeletal Muscle.,J Biol Chem,1968;243:3763-3765.,Earl Wilbur Sutherland,won,Nobel Prize in Physiology or Medicine(,1971,)for their discovery of,concerning the mechanisms of the action of hormones,especially epinephrine,via second messengers(cAMP),Earl Wilbur Sutherland,In 1977,Elliott Ross and Alfred,Gilman,reported a GTP binding protein,which restored the hormone stimulation of purified AC and hormone receptor.,Elliott M.Ross and,Alfred G.Gilman,.,Reconstitution of Catecholamine-Sensitive Adenylate Cyclase Activity:Interaction of Solubilized Components with Receptor-Replete Membranes.,PNAS,Sep 1977;74:3715.,E.M.Ross and,A.G.Gilman,.,Resolution of some components of adenylate cyclase necessary for catalytic activity.,J Biol Chem,Oct 1977;252:6966.,Alfred G.Gilman,&,Martin Rodbell,shared the Nobel Prize in Physiology or Medicine(1994)for their discovery of,G-proteins and the role of these proteins in signal transduction in cells.,G-Protein Coupled Receptor，GPCR,Robert J.Lefkowitz and Brian K.Kobilka,Won 2012 Nobel prize,细胞信号转导研究的必要性,当前细胞生物学研究的三大基本问题之一：基因表达产物主要是大量活性因子与,信号分子,，它们是如何调节细胞最重要的生命活动过程的？,当前细胞基本生命活动研究的四大课题之一:,细胞信号转导的研究,全世界自然科学研究中论文发表最集中的三个领域之一:,细胞信号转导,目前全球研究最热门的五大研究方向之一:,信号转导,目前全球药物开发四分之一集中在：,信号通路上的蛋白激酶,Single cellular organism,respond directly to the environmental,stimulation,Multiple cellular,organism,respond to the environmental stimulation via complex signal transduction system.,When organisms receive environmental stimulus：,Introduction,When an organism receives environmental,stimulus,how does it respond?,Cells must respond to essential signals in their environment to regulate activity of organism.,Stimulus,Target cell,Secrete cell,Secrete signal molecules,Y,Y,Y,Two modes of cell conmmunication:,via,direct contact of adjacent cells,via secretion different kinds of chemicals to regulate the metabolism and function of other cells,Signaling molecules:,chemicals which are able to transduce signal and regulate the cellular activity are,called,signaling molecules.,G,ap junction,（细胞间隙连接）,General mode of information communication between cells,Y,Y,Y,Blood vessel,Signal substances,Y,Receptor,Secretory cell,Target cell,Target cells,Stimulation,How does hormones regulate metabolism of organism,?,How does growth factors or hormones regulate gene expression of organism,?,They need to talk.,How to talk?,Phosphorylase kinase,Phosphorylase kinase,ATP,phosphorylase,phosphorylase,ATP,PPi,Phosphoprotein Phosphohydrolase,Phosphoprotein Phosphohydrolase,H,2,O,PPi,PKA,Inhibitor,a,Inhibitor,b,ATP,Phosphoprotein Phosphohydrolase,PPi,The effects of adrenalin to glycogen metablism,(肾上腺素对糖原代谢的影响),Adrenalin,Receptor,A,drenalin receptor complex,Activate,protein,Activate,AC,ATP,cAMP,PKA,This is an example of the Ras pathway,which results in cell division,.,General steps followed by,transmembrane signal transduction pathway,Signaling molecules,released from specific cells,Signaling molecules reaching target cell,Via blood circulation of diffusion,Binding specifically the,receptor,of target cell,Transferring of signal of receptor,and initiate intracellular system,Biological effects of target cells,Ligand,(First messenger),Receptor,Second messenger,Effecter,proteins,Effecter,proteins,Biological effects of target cells,Signal molecules,Receptors,Signal tranduction pathway,Crosstalk of different signal,tranduction pathways,Signal tranduction pathway and desease,Content of this chapter,Section 1,Signal Molecules,一、,Extracellular signal molecules,Extracellular signal molecules,are small chemical molecules secreted from secreting cells and regulating biologic activity,also called,first messenger,.,Chemical properties,:,*,Proteins and peptides:,Growth factors(EGF,PDGF)、cytokines(TNF、IFN)、insulin et al,*,Amino acids and its derivatives:,Glycine、thyroid、adrenalin et al,*,Steroid hormones:,Glucocorticoid,、sex hormones et al,*,Derivatives of lipids,:,prostaglandin,Gas:,Nitric oxide(NO)、carbon monoxide(CO),Y,Y,Y,Y,Blood vessel,Signal molecule,Y,Receptor,Secret cell,Target cell,Target cell,Stimulation,Target cell,Target cell,Endocrine,Paracrine,一,.Intercellular information substances,*,Definition:,A type of chemical substances secreted by cells can regulate vital activity of target cells.,*,Chemical property:,Such as proteins&peptides,amino acids&its derivation,steroid hormone,fatty acid derivation,and nitric oxide(NO)etc.,（一）,Transmitter,Synaptic signal,Characteristic,：,Secreted from,Neuron,；,via,synaptic cleft,reaching next neuron；,Short action time.,Example：,acetylcholine(Ach)、noradrenaline(NA),（二）,Endocrine hormones,Endocrine signal（hormones）,Characteristic,：,secretion from autocrine cells;,via blood circulation to get to,the target cell；,Long action time,Example：,Insulin、thyroid、adrenalin et al,（内分泌）,（三）,Paracrine signal,（旁分泌）,Characteristic,：,Secreted from normal cells；,via diffusion to reach target cell.,Shorter action time,Example：,Growth factors、cytokines et al,（四）,Gas signal,Nitric oxide(NO),Others,Some signal molecules can function for same type of cells or even regulate autocrine cells and are called,autocrine signal（自分泌）.,Some signal molecules can be transmitted among different bodies.,Cellular functional pathway influenced by different kinds of signal molecules,Types,Signal molecules,Receptor,Biologic effects,Neuron transmitter,acetylcholine,、,glutarate;GA,、,-,aminobutyrate,Membrane receptor,Influence ion channel,Growth factors,IGF-1、EGF、PDGF,Membrane receptor,Leading to phosphorylation or dephosphorylation of enzyme or functional proteins and change to change the metabolism and gene expression of cells,Hormones,Hormones of proteins、polypeptides and derivatives of amino acids,Steroid、thyroid hormone,Membrane receptor,Intracellular receptors,The same as above,Regulate transcription,Vitamines,Vitamine A、Vitamine D,Intracellular receptors,The same as above,二、,I,ntracellular signal molecules：,I,ntracellular signal molecules：,Chemicals which is able to respond to the signal and transduce signal within cells.,Chemical properties,Inorganic ion：such as,Ca,2+,Derivatives of lipid：,DAG,、,Cer,Derivatives of carbonhydrates：,IP,3,Nucleotides：,cAMP、cGMP,Signal protein molecules,Third messenger,Is responsible for transmitting intranuclear signal，they are normally called DNA binding protein.It is a type of DNA binding protein which can specific DNA sequence and regulate gene expression,eg proteins encoded by immediate-early genes.,Intracellular small molecules which can tranduce signal information，such as Ca,2+,、,DAG、IP,3,、,Cer、cAMP、cGMP st al.,Secondary messenger,Section 2,Receptor,Each cell in a multi-cellular organism is exposed to many-perhaps hundreds-of different signals from its environment.These signals must be received either at the cell surface or in the cytoplasm by specialized proteins that form a transmission system for signal transduction.,The biologic active molecules which can bind to receptor specifically and called,ligands.,Receptors,They are special proteins which are able to recognize extracellular signal molecules,transmit the signal into cells and lead to biological effect.During the process the signal is enlarged and transmitted correctly.,One type of DNA-binding protein located in cytosol or neucleus.,Most of them are mosaic glycoproteins,located on plasma membrane of cell surface.,Intracellular receptor:,Membrane receptor:,According to the location within the cell:,Classification、structure and function,of receptor,Located at the cell membrane.Most of them are intercalating glyproteins.It is classifiedinto three types：,ion channel,，,G-protein coupled receptors,and single transmembrane receptors.,（一）,Membrane receptor,1.Circular receptor Ligand dependent ion channel receptor,Acetylcholine receptor;,Ach/toxin,Binding site,Ach/toxin,Binding site,Ach/toxin,Binding site,Ach/toxin,Binding site,Inside of cell,Outside of cell,2.G-protein coupled receptors(GPCRs),(S,erpentine receptor),G蛋白偶联受体的结构,矩型代表,-螺旋，N端被糖基化，C端的半胱氨酸被棕榈酰化。,G-protein coupled receptors,Special feature of the structure of the receptor：,N terminal contain multiple,glycosylation sites,*,Contain high conservative cysteine residue，which play important role in maintaining the structure of receptor.,*,The second and third loop are the G protein coupled region.,Typically,G proteins are,heterotrimers,consisting of,-(45 to 47 kD),-(35 kD),and,-(7 to 9 kD)subunits.,Binding of hormone to its receptor causes a conformational change that induces the receptor to catalyze a,replacement,of,GDP,by,GTP,on G,.The,G,(GTP)complex,dissociates from,G,and binds to,adenylyl cyclase,stimulating cAMP synthesis.,G-protein：,G,uanylate binding protein,is a kind of GTP or GDP binding protein which is located at the out side of the cytoplasm membrane and consists of,、,subunits.,Two kinds of conformation:,active-and inactive-types,Inactive type,Active type,Hormone receptor acts through G proteins.These G proteins are coupled to several pathways,including,adenylyl cyclase,guanylyl cyclase,phospholipases A and,C,Ca2+,K+channels,.,R,H,AC,GDP,GTP,Adenylcyclase,AC,ATP,cAMP,protein,G-protein in signal transduction,Types,subunit function,G,s,s,i,Inhibites AC,q,q,Activates LPL-C specific,PLC,o,*,o,Regulator G-protein of,ion channel in brain,Transmitter,a,Activates visual sense,protein,Activates AC,Signal transduction pathway mediated by this kind of receptors,Hormone,Receptor,protein,Enzyme,Second messager,Protein kinase,Enzymes and functional proteins,Biological effect,Cys-rich region,TPK,EGF-R,IGF-I-R,PDGF-R,FGF-R,Kinase-inserted region,Singal transmembrane,-helix receptor,：,Membrane,Ig-like sequence,Single transmembrane,helix,Receptor contain TPK domain,EGF:表皮生长因子 IGF-1:胰岛素样生长因子,PDGF:血小板衍生生长因子 FGF:成纤维细胞生长因子,After ligand binding,the receptor have tyrosine protein kinase activity such as,insulin receptor,IGF-R,epidermal growth factor receptor,EGF-R.,After ligand binding,can couple with tyrosine protein kinase and have tyrosine protein kinase activity growth factor.,Non tyrosine protein kinase receptor,Receptor tyrosine protein kinase,A,utophosphorylation,When ligand bind to receptor,，,the catalytic receptors dimerize，dimerized tyrosine protein kinase(TPK)are activated，each of them phosphorylates tyrosine of the other monomer,this process is called autophosphorylation.,This type of receptor is related with cell proliferation,differentiation and tumorgenesis,Mechanism of EGF-receptor:,*,A single t,ransmembrane domain,consists of 2226 amino-acid residue and form hydrophobic-helix.,*,E,xtracellular region is ligand binding sites.,*,A cytoplasmic tyrosine kinase domain o,r called SH,1,Src homology 1 domain，located at C terminal，including two functional region:ATP and substrate binding region.,Structural of receptor：,Type and type of TGFreceptor,Ligand,Kinase domain,Kinase domain,4.Receptor with guanylate cyclase activity,胞外,胞内,Membrane receptor,Soluble receptor,PKH,GC,GC,Structure of,receptor with guanylate cyclase activity,PKH：Kinase like domain,GC：guanylate cyclase domain,guanylate cyclase,No tyrosine kinase activity but contian consevative,-,helix structure,which can bind,to tyrosine kinase JAK,（,Janus kinase family,）.After binding,the signal can be transmitted via,signal transducer and activator of transcription,，(,STAT,).,（,3,）C,ytokine receptor,(,细胞因子受体,),Structure of,intracellular,receptor,（二）,Intracellular receptor,They are DNA binding protein and located at cytoplasm or nucleus.,High variable region,N terminal，transcriptional,activation activity,DNA binding region,Contain zinc structure,Hormone binding domain,C terminal，heat shock，dimerization of receptors，Transcriptional activation,Hinge region,Binding site of hormones,Structure of,intracellular,receptor,High variable region,DNA binding region,Hormone binding domain,Hinge region,Corresponding ligand,Steroid hormones、thyroid and retinoic acid,Function,serving as trans-acting factors.After ligand binding,they are able to bind to the cis-acting elements to regulate gene regulation.,肿瘤坏死因子,（,tumor necrosis factor,，,TNF,）：受体家族既不含组成性蛋白激酶区域，也不结合胞浆的酪氨酸蛋白激酶或,G,蛋白，而是藉其胞浆段的趋同性区域（,homotropic domain,），,结合连接分子,（,adaptors,）进行信号转导。如死亡受体介导的死亡信号转导。,（,5,）,T,umor necrosis factor,(,TNF,)receptor family,High specificity,High affinity,Saturability,Specific working model,Reversible,配体浓度,受体饱和度（）,Binding curve of ligand receptor binding,Concentration of ligand,二,.Characteristic of receptor action,三、,Regulation of activity of receptor,Phosphorylation and dephosphorylation,Influence the metabolism of,membrane phospholipid,Enzymatic hydrolysis,Regulation of G-protein,Section 2,Signal Transduction Pathway,一、,Signal pathway mediated by transmembrane receptors,cAMP-protein kinase pathway,Ca,2+,-dependent protein kinase pathway,cGMP-protein kinase pathway,Tyrosine protein kinase,pathway,Nuclear,pathway,TGF-pathway,（一）,cAMP protein kinase pathway,Composition,Extracellular signal molecules，receptors such as,-,Adrenergic(肾上腺素)receptor,，G prot