拟南芥Annexin5的克隆及其功能分析
摘要
Annexin家族是一类进化上保守的钙依赖性磷脂、膜结合蛋白,其成员广泛分布在各种真核生物的不同细胞中。大量研究表明,膜联蛋白参与许多重要的细胞活动,例如膜重组、膜运输、分泌活动、囊泡运输等,但是大多数膜联蛋白的确切生理功能仍然是未知之谜。在高等植物中鲜有对于Annexin5基因的研究或报道,因此,本研究项目拟采用分子生物学和细胞生物学方法,首次尝试研究拟南芥花粉特异表达的膜联蛋白家族成员——Annexin5,得到以下结果：
1.克隆得到Annexin5的全长cDNA序列以及Annexin5的钙离子结合位点突变、PIP2结合位点突变、缺失突变等突变序列,并将其插入植物表达载体pCAMBIA1300中与GFP基因融合,本实验中我们一共构建了两套植物表达载体,分别用花粉特异启动子LAT52和ZM13启动表达目的基因。
2.利用农杆菌介导花序浸泡法转化野生型拟南芥,用潮霉素进行筛选,因其带有GFP荧光定位,也可以用荧光显微镜进一步检测目的基因是否转入拟南芥中,目前已经获得全部的转基因植株。
3.筛选稳定表达的转基因植株并观察表型,目前己获得1300-lat52-GFP和1300-lat52-GFP-Ann5的纯合转基因植株,观察这两种植株中的Annexin5过表达对拟南芥花粉萌发以及花粉管生长的影响,旨在为下一步探讨膜联蛋白参与调控花粉管极性生长的分子机制打下基础。
本实验为以后进一步揭示膜联蛋白家族成员在花粉管极性生长过程中作为“枢纽”蛋白衔接膜、微丝骨架和钙离子之间相互作用的分子机制研究打下了基础,具有重要的理论价值和应用前景。
Annexin are Ca2+ and phospholipid binding proteins forming an evolutionary conserved multigene family, with members of the family being expressed throughout animal and plant kingdoms. It is generally assumed that Annexins are implicated in several processes related to membranes, including regulation of membrane organisation, membrane trafficking, secretion and vesicular trafficking.But we are still far from a comprehensive description of Annexin function in cell processes. There is hardly any research or report about the Annexin5 of higher plant. Therefore, this research project is to use molecular biology and cell biology methods, the first attempt to study Arabidopsis pollen-specific expression of the membrane protein family members-Annexin5. Results are as follows:
1. In order to investigate the gene function of Annexin5, the full length of cDNA was cloned and inserted into the plant expression vector of pCAMBIA1300,then fused to GFP gene. We also constructed a set of plant expression vector include mutational or deleted position.for example, mutational in Ca2+ binging position、mutational in PIP2 binging position. In this study,We use pollen special promoter LAT52 and ZM13.
2. The recombination vector was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens through the floral dip method. Filtered by hygromyein, meanwhile, fluoroscopic examination proved that the plant expression vectors had been integrated into arabidopsis genome. Now we have got all the transgenic plants.
3. Screening transgenic plants stably expressed and observed its phenotype, now we have obtained homozygotes of 1300-lat52-GFP and 1300-lat52-GFP-Ann5. We confirmed that whether the overexpression of Annexin5 has influences on the pollen germination、pollen tube growth and actin cytoskeleton. By observe the effect of Annexin5 on Arabidopsis thaliana pollen tube growth, we wanted to reveal the molecular mechanism of membrane protein involved in pollen germination and pollen tube growth.
This study laid a foundation for revealed the molecular mechanism of members of the annexin family, Which act as Hub protein play an important role in the interactions among membrane, actin cytoskeleton and calcium ions during polar growth of pollen tube. has important theoretical value and application prospect.
1.克隆得到Annexin5的全长cDNA序列以及Annexin5的钙离子结合位点突变、PIP2结合位点突变、缺失突变等突变序列,并将其插入植物表达载体pCAMBIA1300中与GFP基因融合,本实验中我们一共构建了两套植物表达载体,分别用花粉特异启动子LAT52和ZM13启动表达目的基因。
2.利用农杆菌介导花序浸泡法转化野生型拟南芥,用潮霉素进行筛选,因其带有GFP荧光定位,也可以用荧光显微镜进一步检测目的基因是否转入拟南芥中,目前已经获得全部的转基因植株。
3.筛选稳定表达的转基因植株并观察表型,目前己获得1300-lat52-GFP和1300-lat52-GFP-Ann5的纯合转基因植株,观察这两种植株中的Annexin5过表达对拟南芥花粉萌发以及花粉管生长的影响,旨在为下一步探讨膜联蛋白参与调控花粉管极性生长的分子机制打下基础。
本实验为以后进一步揭示膜联蛋白家族成员在花粉管极性生长过程中作为“枢纽”蛋白衔接膜、微丝骨架和钙离子之间相互作用的分子机制研究打下了基础,具有重要的理论价值和应用前景。
Annexin are Ca2+ and phospholipid binding proteins forming an evolutionary conserved multigene family, with members of the family being expressed throughout animal and plant kingdoms. It is generally assumed that Annexins are implicated in several processes related to membranes, including regulation of membrane organisation, membrane trafficking, secretion and vesicular trafficking.But we are still far from a comprehensive description of Annexin function in cell processes. There is hardly any research or report about the Annexin5 of higher plant. Therefore, this research project is to use molecular biology and cell biology methods, the first attempt to study Arabidopsis pollen-specific expression of the membrane protein family members-Annexin5. Results are as follows:
1. In order to investigate the gene function of Annexin5, the full length of cDNA was cloned and inserted into the plant expression vector of pCAMBIA1300,then fused to GFP gene. We also constructed a set of plant expression vector include mutational or deleted position.for example, mutational in Ca2+ binging position、mutational in PIP2 binging position. In this study,We use pollen special promoter LAT52 and ZM13.
2. The recombination vector was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens through the floral dip method. Filtered by hygromyein, meanwhile, fluoroscopic examination proved that the plant expression vectors had been integrated into arabidopsis genome. Now we have got all the transgenic plants.
3. Screening transgenic plants stably expressed and observed its phenotype, now we have obtained homozygotes of 1300-lat52-GFP and 1300-lat52-GFP-Ann5. We confirmed that whether the overexpression of Annexin5 has influences on the pollen germination、pollen tube growth and actin cytoskeleton. By observe the effect of Annexin5 on Arabidopsis thaliana pollen tube growth, we wanted to reveal the molecular mechanism of membrane protein involved in pollen germination and pollen tube growth.
This study laid a foundation for revealed the molecular mechanism of members of the annexin family, Which act as Hub protein play an important role in the interactions among membrane, actin cytoskeleton and calcium ions during polar growth of pollen tube. has important theoretical value and application prospect.
引文
Alvarez-Martinez, M.T., Mani, J.C., Porte, F., Faivre-Sarrailh, C., Liautard, J.P., and Sri-Widada, J. (1996). Characterization of the interaction between Annexin I and profilin. Eur J Biochem, 238:777-784.
Alvarez-Martinez, M.T., Porte, F., Liautard, J.P., and Sri-Widada, J. (1997). Effects of profilin-Annexin 1 association on some properties of both profiling and Annexin Ⅰ: modification of the inhibitory activity of profilin on actin polymerization and inhibition of the self-association of Annexin I and its interactions with liposomes. Biochim Biophys Acta,1339:331-340.
Babiychuk, EB., Monastyrskaya, K., Burkhard, F.C., Wray, S., Draeger, A.(2002) Modulating signaling events in smooth muscle:cleavage of Annexin2 abolishes its binding to lipid rafts. FASEB J 16:1177-1184
Babiychuk, EB., Palstra, RJ., Schaller, J., Kampfer,U., Draeger,A.(1999) Annexin VI participates in the formation of a reversible, membrane-cytoskeleton complex in smooth muscle cells. J Biol Chem 274:35191-35195 1405-1418.
Bianchi, MW., Damerval, C., Vartanian, N. (2002)Identification of proteins regulated bycross-talk between drought and hormone pathways in Arabidopsis wild-type and auxin-insensitive mutants, axrl and axr2.Funct Plant Biol 29:51-61.
Blackbourn, H.D., Barker, P.J., Huskisson, N.S., and Battey, N.H. (1992). Properties and partial protein sequence of plant Annexins. Plant Physiol,99:864-871.
Breton, G., Vazquez-Tello, A., Danyluk, J., and Sarhan, F. (2000). Two novel intrinsic Annexins accumulate in wheat membranes in response to low temperature. Plant Cell Physiol,41: 177-184.
Buzan, JM., Frieden,C.(1996) Yeast actin:polymerization kinetic studies of wild type and a poorly polymerizing mutant. Proc Natl Acad Sci USA 93:91-95
Campanoni, P., and Blatt, M.R. (2007). Membrane trafficking and polar growth in root hairs and pollen tubes. J Exp Bot,58:65-74.
Caroline, M.C., Stephanie, J.G., and Dianna, J.B. (1996). Tomato Annexins p34 and p35 bind to F-actin and display nucleotide phosphodiesterase activity inhibited by phospholipids binding. Plant Cell,8:333-342.
Cheung, A.Y., and Wu, H.M. (2001). Pollen tube guidance:Right on target. Science,293: 1441-1442.
Clark, GB., Dauwalder, M., Roux,SJ. (1998)Immunological and biochemical evidence for nuclear localization of Annexin in peas. Plant Physiol Biochem 36:621-627
Clark, G.B., Thompson, G.J., and Roux, S.J. (2001). Signal transduction mechanisms in plants: an overview. Curr Sci,80:170-177.
Creutz,CE., Pazoles,CJ., Pollard,HB. (1978) Identification and purification of an adrenal medullary protein (synexin) that causes calcium-dependent aggregation of isolated chromaffin granules. J Biol Chem 253:2858-2866.
Cardenas, L., Lovy-Wheeler, A., Kunkel, J.G., and Hepler, P.K. (2008) Pollen tube growth oscillations and intracellular calcium levels are reversibly modulated by actin polymerization. Plant physiol, Published online on February 8,2008, as DOI:10.1104/pp.107.113035.
D. Konopka-Postupolska(2007).Annexins:putative linkers in dynamic membrane- cytoskeleton interactions in plant cells. Protoplasma 230:203-215
Dorota,K.P., Greg,Clark.,(2009).The Role of Annexin 1 in Drought Stress in Arabidopsis 1[W].Plant Physiology,Vol.150, pp.1394-1410,
Filipenko, N.R., and Waisman, D.M. (2001). The C-terminus of Annexin Ⅱ mediates binding to F-actin. J Biol Chem,276:5310-5315.
Funakoshi,T.,Hendrickson,E.,McMullen,A., (1987), Primary structure of human placental anticoagulant protein.Biochemistry,26(25):8087-8092
Geitmann, A., and Emons, A.M. (2000). The cytoskeleton in plant and fungal cell tip growth. J Microsc,198:218-245.
Gerke, V., and Moss, S.E. (2002). Annexins:from structure to function. Physiol Rev,82: 331-371.
Gidrol,X., Sabelli,PA., Fern,YS., Kush,AK.(1996) Annexin-like protein from Arabidopsis thaliana rescues delta oxyR mutant of Escherichia coli from H2O2 stress. Proc Natl Acad Sci USA 93:11268-11273
Hofmann, A., Proust, J., Dorowski, A., Schantz, R., and Huber, R. (2000). Annexin 24 from Capsicum annuum. X-ray structure and biochemical characterization. J Biol Chem,275: 8072-8082.
Hoshino,D., Hayashi,A., Temmei,Y., Kanzawa,N., Tsuchiya,T. (2004) Biochemical and immunohistochemical characterization of Mimosa Annexin. Planta 219:867-875
Huber,R., Berendes,R., Burger,A. (1992),Crystal and molecular structure of human AnnexinV after refinement.Implications for structure,membrane binding and ion channel formation of the Annexin family of proteins. J.Mol.Biol,223(3):683-704
Janicke,RU., Porter,AG.,sh,A.(1998) A novel Arabidopsis thaliana protein protects tumor cells from tumor necrosis factor-induced apoptosis. Biochim Biophys Acta 1402:70-78
Janmey,PA., Stossel,TP.(1987) Modulation of gelsolin function by polyphosphoinositol (4,5)-bisphosphate. Nature 325:362-364
Khanna,NC., Helwig,ED., Ikebuchi,NW., Fitzpatrick,S., Bajwa,R.,Waisman,DM.(1990) Purification and characterization of Annexin proteins from bovine lung. Biochemistry 29: 4852-4862
Kim,E., Miller.CJ., Reisler,E.(1996) Polymerization and in vitro motility properties of yeast actin:a comparison with rabbit skeletal alpha actin. Biochemistry 35:16566-16572
Konopka-Postupolska, D. (2007). Annexins:putative linkers in dynamic membrane-cytoskeleton interactions in plant cells. Protoplasma,230:203-215.
Kovacs,I., Ayaydin,F., Oberschall,A., Ipacs,I., Bottka,S., Dudits,D., Toth,EC.(1998) Immunolocalization of a novel Annexin-like protein encoded by a stress and abscisic acid responsive gene in alfalfa. Plant J 15:185-197
Kreps,JA., Wu YJ, Chang HS, Zhu T, Wang X, Harper JF (2002) Transcriptome changes for Arabidopsis in response to salt, osmotic, and cold stress. Plant Physiol 130:2129-2141
Kush,A., Sabapathy,K.(2001) Oxy5, a novel protein from Arabidopsis thaliana protects mammalian cells from oxidative stress. Int J Biochem Cell Biol 33:591-60
Lee, DBN., Jamgotchian,N., Allen,SG., Kan,FWK., Hale,IL. (2004) Annexin A2 heterotetramer: role in tight junction assembly. Am J Physiol Renal Physiol 287:F481-F491
Lee, S., Lee,EJ., Yang, EJ., Lee,JE., Park,AR., Song,WH., Park,OK.(2004) Proteomic identification of Annexins, calcium-dependent membrane binding proteins that mediate osmotic stress and abscisic acid signal transduction in Arabidopsis. Plant Cell 16: 1378-1391
Messerli, M.A., Danuser, G., and Robinson, R.R. (1999). Pulsatile influxes of H+, K+ and Ca2+ lag growth pulses of Lilium longiflorum pollen tubes. J cell Sci,112:1497-1509.
Mortimer,JC., Laohavisit, A., Macpherson, N.,Webb,A.,Brownlee,C., Battey,NH., Davies,JM.(2008) Annexins:multifunctional components of growth and adaptation. J Exp Bot 59:533-544
Proust,J., Houlne,G., Schantz,ML., Schantz,R. (1996) Characterization an gene expression of an Annexin during fruit development in Capsicuannum. FEBS Lett 383:208-212
Ren, H.Y., and Xiang, Y. (2007). The function of actin-binding proteins in pollen tube growth. Protoplasm,230:171-182.
Rothman, JE. (1994) Mechanisms of intracellular transport. Nature 372:55-63
Santoni,V., Rouquie,D., Doumas, P., Mansion,M., Boutry,M., Degand H,D. upree,P., Packman,L., Sherrier,J., Prime,T., Bauw,G., Posada,E., Rouze,P., Dehais,P., Sahnoun,I., Barlier, I.,Rossignol,M.(1998) Use of a proteome strategy for tagging proteins present at the plasma membrane Plant J 16:622-643
Schlaepfer, DD., Haigler,HT. (1987) Characterization of Ca2+-dependent phospholipid binding and phosphorylation of lipocortin I. J BiolChem 262:6931-6937
Seigneurin-Berny, D., Rolland,N., Dome,A-J., Joyard,J.(2000) Sulfolipid is a potential candidate for Annexin binding to the outer surface of chloroplast. Biochem Biophys Res Commun 272:519-524
Shin,H., Kudlicka,K., Brown,RM.(1995) A biochemical study on glucan synthesis in the cotton fiber (abstract no.298). Plant Physiol 108:S-68
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Smallwood,M., Keen,JN., Bowles,DJ.(1990) Purification and partial sequence analysis of plant Annexins. Biochem J 270:157-161
Sopkova,J.,Renouard,M.,Lewit-Bentley,A., (1993)The crystal structure of a new high-calcium form of Annexin V.J.Mol.Biol,1993,234(3):816-825
Swairjo,MA., Concha,NO., Kaetzel,MA., Dedman,JR., Seaton,BA.(1995)Ca2+bridging mechanism and phospholipids head group recognition in the membrane-binding protein Annexin V. Nat Struct Biol 2:968-974
Thonat,C., Mathieu,C., Crevecoeur,M., Penel,C., Gaspar T, Boyer N(1997) Effects of a mechanical stimulation on localization of Annexin-like proteins in Bryonia dioica internodes. Plant Phvsiol 114:981-988
Traverso,V., Morris,JE, Flower,RJ., Buckingham, J.(1998) Lipocortin 1(Annexin 1) in patches associated with the membrane of a lung adenocarcinoma cell line and in the cell cytoplasm. J Cell Sci 111:1405-1418
Tzima,E., Trotter,PJ., Orchard,MA., Walker,JH. (2000) AnnexinV relocates to the platelet cytoskeleton upon activation and binds to a specific isoform of actin. Eur J Biochem 267: 4720-4730
Ursula,R.,Volker, G. (2004).Annexins - unique membrane binding proteins with diverse functions. Journal of Cell Science 117,2631-2639
Volker,G. Stephen, E. Moss.(2002). Annexins:From Structure to Function. Physiol Rev 82:331-371,
Xiang, Y., Huang, X., Wang, T., Zhang, Y., Liu, Q., Hussey, P.J., and Ren, H.Y. (2007). ACTIN BINDING PROTEIN29 from Lilium pollen plays an important role in dynamic actin remodeling. Plant Cell,19:1930-1946.
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李欣2006《膜联蛋白A5与宫颈粘膜上皮癌变的关系》,解剖学杂志,29(4)：434-435
张毅,陈蕊雯,孙树汉等.(2002)一种新的钙依赖性磷脂结合蛋白Annexin32对血凝及
血栓形成的影响.中国科学(C辑).32(2)：172-176.
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Alvarez-Martinez, M.T., Porte, F., Liautard, J.P., and Sri-Widada, J. (1997). Effects of profilin-Annexin 1 association on some properties of both profiling and Annexin Ⅰ: modification of the inhibitory activity of profilin on actin polymerization and inhibition of the self-association of Annexin I and its interactions with liposomes. Biochim Biophys Acta,1339:331-340.
Babiychuk, EB., Monastyrskaya, K., Burkhard, F.C., Wray, S., Draeger, A.(2002) Modulating signaling events in smooth muscle:cleavage of Annexin2 abolishes its binding to lipid rafts. FASEB J 16:1177-1184
Babiychuk, EB., Palstra, RJ., Schaller, J., Kampfer,U., Draeger,A.(1999) Annexin VI participates in the formation of a reversible, membrane-cytoskeleton complex in smooth muscle cells. J Biol Chem 274:35191-35195 1405-1418.
Bianchi, MW., Damerval, C., Vartanian, N. (2002)Identification of proteins regulated bycross-talk between drought and hormone pathways in Arabidopsis wild-type and auxin-insensitive mutants, axrl and axr2.Funct Plant Biol 29:51-61.
Blackbourn, H.D., Barker, P.J., Huskisson, N.S., and Battey, N.H. (1992). Properties and partial protein sequence of plant Annexins. Plant Physiol,99:864-871.
Breton, G., Vazquez-Tello, A., Danyluk, J., and Sarhan, F. (2000). Two novel intrinsic Annexins accumulate in wheat membranes in response to low temperature. Plant Cell Physiol,41: 177-184.
Buzan, JM., Frieden,C.(1996) Yeast actin:polymerization kinetic studies of wild type and a poorly polymerizing mutant. Proc Natl Acad Sci USA 93:91-95
Campanoni, P., and Blatt, M.R. (2007). Membrane trafficking and polar growth in root hairs and pollen tubes. J Exp Bot,58:65-74.
Caroline, M.C., Stephanie, J.G., and Dianna, J.B. (1996). Tomato Annexins p34 and p35 bind to F-actin and display nucleotide phosphodiesterase activity inhibited by phospholipids binding. Plant Cell,8:333-342.
Cheung, A.Y., and Wu, H.M. (2001). Pollen tube guidance:Right on target. Science,293: 1441-1442.
Clark, GB., Dauwalder, M., Roux,SJ. (1998)Immunological and biochemical evidence for nuclear localization of Annexin in peas. Plant Physiol Biochem 36:621-627
Clark, G.B., Thompson, G.J., and Roux, S.J. (2001). Signal transduction mechanisms in plants: an overview. Curr Sci,80:170-177.
Creutz,CE., Pazoles,CJ., Pollard,HB. (1978) Identification and purification of an adrenal medullary protein (synexin) that causes calcium-dependent aggregation of isolated chromaffin granules. J Biol Chem 253:2858-2866.
Cardenas, L., Lovy-Wheeler, A., Kunkel, J.G., and Hepler, P.K. (2008) Pollen tube growth oscillations and intracellular calcium levels are reversibly modulated by actin polymerization. Plant physiol, Published online on February 8,2008, as DOI:10.1104/pp.107.113035.
D. Konopka-Postupolska(2007).Annexins:putative linkers in dynamic membrane- cytoskeleton interactions in plant cells. Protoplasma 230:203-215
Dorota,K.P., Greg,Clark.,(2009).The Role of Annexin 1 in Drought Stress in Arabidopsis 1[W].Plant Physiology,Vol.150, pp.1394-1410,
Filipenko, N.R., and Waisman, D.M. (2001). The C-terminus of Annexin Ⅱ mediates binding to F-actin. J Biol Chem,276:5310-5315.
Funakoshi,T.,Hendrickson,E.,McMullen,A., (1987), Primary structure of human placental anticoagulant protein.Biochemistry,26(25):8087-8092
Geitmann, A., and Emons, A.M. (2000). The cytoskeleton in plant and fungal cell tip growth. J Microsc,198:218-245.
Gerke, V., and Moss, S.E. (2002). Annexins:from structure to function. Physiol Rev,82: 331-371.
Gidrol,X., Sabelli,PA., Fern,YS., Kush,AK.(1996) Annexin-like protein from Arabidopsis thaliana rescues delta oxyR mutant of Escherichia coli from H2O2 stress. Proc Natl Acad Sci USA 93:11268-11273
Hofmann, A., Proust, J., Dorowski, A., Schantz, R., and Huber, R. (2000). Annexin 24 from Capsicum annuum. X-ray structure and biochemical characterization. J Biol Chem,275: 8072-8082.
Hoshino,D., Hayashi,A., Temmei,Y., Kanzawa,N., Tsuchiya,T. (2004) Biochemical and immunohistochemical characterization of Mimosa Annexin. Planta 219:867-875
Huber,R., Berendes,R., Burger,A. (1992),Crystal and molecular structure of human AnnexinV after refinement.Implications for structure,membrane binding and ion channel formation of the Annexin family of proteins. J.Mol.Biol,223(3):683-704
Janicke,RU., Porter,AG.,sh,A.(1998) A novel Arabidopsis thaliana protein protects tumor cells from tumor necrosis factor-induced apoptosis. Biochim Biophys Acta 1402:70-78
Janmey,PA., Stossel,TP.(1987) Modulation of gelsolin function by polyphosphoinositol (4,5)-bisphosphate. Nature 325:362-364
Khanna,NC., Helwig,ED., Ikebuchi,NW., Fitzpatrick,S., Bajwa,R.,Waisman,DM.(1990) Purification and characterization of Annexin proteins from bovine lung. Biochemistry 29: 4852-4862
Kim,E., Miller.CJ., Reisler,E.(1996) Polymerization and in vitro motility properties of yeast actin:a comparison with rabbit skeletal alpha actin. Biochemistry 35:16566-16572
Konopka-Postupolska, D. (2007). Annexins:putative linkers in dynamic membrane-cytoskeleton interactions in plant cells. Protoplasma,230:203-215.
Kovacs,I., Ayaydin,F., Oberschall,A., Ipacs,I., Bottka,S., Dudits,D., Toth,EC.(1998) Immunolocalization of a novel Annexin-like protein encoded by a stress and abscisic acid responsive gene in alfalfa. Plant J 15:185-197
Kreps,JA., Wu YJ, Chang HS, Zhu T, Wang X, Harper JF (2002) Transcriptome changes for Arabidopsis in response to salt, osmotic, and cold stress. Plant Physiol 130:2129-2141
Kush,A., Sabapathy,K.(2001) Oxy5, a novel protein from Arabidopsis thaliana protects mammalian cells from oxidative stress. Int J Biochem Cell Biol 33:591-60
Lee, DBN., Jamgotchian,N., Allen,SG., Kan,FWK., Hale,IL. (2004) Annexin A2 heterotetramer: role in tight junction assembly. Am J Physiol Renal Physiol 287:F481-F491
Lee, S., Lee,EJ., Yang, EJ., Lee,JE., Park,AR., Song,WH., Park,OK.(2004) Proteomic identification of Annexins, calcium-dependent membrane binding proteins that mediate osmotic stress and abscisic acid signal transduction in Arabidopsis. Plant Cell 16: 1378-1391
Messerli, M.A., Danuser, G., and Robinson, R.R. (1999). Pulsatile influxes of H+, K+ and Ca2+ lag growth pulses of Lilium longiflorum pollen tubes. J cell Sci,112:1497-1509.
Mortimer,JC., Laohavisit, A., Macpherson, N.,Webb,A.,Brownlee,C., Battey,NH., Davies,JM.(2008) Annexins:multifunctional components of growth and adaptation. J Exp Bot 59:533-544
Proust,J., Houlne,G., Schantz,ML., Schantz,R. (1996) Characterization an gene expression of an Annexin during fruit development in Capsicuannum. FEBS Lett 383:208-212
Ren, H.Y., and Xiang, Y. (2007). The function of actin-binding proteins in pollen tube growth. Protoplasm,230:171-182.
Rothman, JE. (1994) Mechanisms of intracellular transport. Nature 372:55-63
Santoni,V., Rouquie,D., Doumas, P., Mansion,M., Boutry,M., Degand H,D. upree,P., Packman,L., Sherrier,J., Prime,T., Bauw,G., Posada,E., Rouze,P., Dehais,P., Sahnoun,I., Barlier, I.,Rossignol,M.(1998) Use of a proteome strategy for tagging proteins present at the plasma membrane Plant J 16:622-643
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