Two-dimensional zymography in detection of proteolytic enzymes in wheat leaves

• Autorzy: Grudkowska M. , Lisik P. , Rybka K.
• Języki publikacji: EN

Abstrakty

EN

Proteolytic enzymes in wheat leaves were studied using zymographic detection of enzyme activities on one-way (1D) SDS-polyacrylamide gels and two-dimensional (2D) ones, on which protein samples were isoelectrofocused prior to PAGE separation. Gelatin of concentration 0.1 %, copolymerized into SDS-PAGE gels, digested by active proteinases enabled detection of those enzymes. On 1D gels, seven bands were seen and assigned to particular families through the use of specific inhibitors. Metalloproteinases inhibited by 20 mM EDTA were detected as 150 kDa band; aspartic proteinases were assigned to 115–118 kDa double band by using 25 mM pepstatin; 10 mM phenylmethylsulfonyl fluoride used for detection of serine proteinases pointed to band of 70 kDa and finally due to 10 μM E-64 inhibitor, cysteine proteinases of 37 and 40 kDa were detected. On 2D gels, additional separation according to protein isoelectric points enabled detection of proteinase isoforms. In the range of 4.5–6 pI, six metalloproteinases as well as ten aspartic proteinases were visible, ten serine- isoforms of pI 4.5–6.8 and four cysteine proteinases of 4.5–5.0 pI were found. Presented results were detected as reproducible results observed at least in four independent biological replications.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2013
• Tom: 35
• Numer: 12
• Opis fizyczny: p.3477-3482,fig.,ref.

Twórcy

autor

Grudkowska M.

• Department of Biochemistry, Faculty of Agriculture and Biology, Warsaw University of Life Sciences, SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland

autor

Lisik P.

• Department of Biochemistry, Faculty of Agriculture and Biology, Warsaw University of Life Sciences, SGGW, Nowoursynowska 159, 02-776 Warsaw, Poland

autor

Rybka K.

• Plant Physiology and Biochemistry Department, Plant Breeding and Acclimatization Institute–National Research Institute, Radziko´w, 05-870 Błonie, Poland

Bibliografia

• Afshar-Mohammadian M, Rahimi-Koldeh J, Sajedi R (2011) The comparison of protease activity and total protein in three cultivars of kiwifruit of Northern Iran during fruit development. Acta Physiol Plant 33:343–348. doi: 10.1007/s11738-010-0553-3
• Cazenave C, Toulme JJ (2001) Gel renaturation assay for ribonucleases. In: Nicholson AW (ed) Methods of enzymology, vol 341. Academic Press, New York, pp 126–141. doi: 10.1016/S0076-6879(01)41149-9
• Chi W, Sun X, Zhang L (2012) The roles of chloroplast proteases in the biogenesis and maintenance of photosystem II. Biochim Biophys Acta Bioenerg 1817:239–246. doi: 10.1016/j.bbabio.2011.05.014
• Chichkova NV, Tuzhikov AI, Taliansky M, Vartapetian AB (2012) Plant phytaspases and animal caspases: structurally unrelated death proteases with a common role and specificity. Physiol Plant 145:77–84. doi: 10.1111/j.1399-3054.2011.01560.x
• Cho CW, Chung E, Kim K et al (2009) Plasma membrane localization of soybean matrix metalloproteinase differentially induced by senescence and abiotic stress. Biol Plant 53:461–467. doi: 10.1007/s10535-009-0086-z
• George S, Johnson J (2010) In Situ zymography. In: Clark IM (ed) Matrix metalloproteinase protocols, vol 622. Humana Press, New Jersey, pp 271–277. doi: 10.1007/978-1-60327-299-5_17
• Grbić V (2003) SAG2 and SAG12 protein expression in senescing Arabidopsis plants. Physiol Plant 119:263–271. doi: 10.1034/j.1399-3054.2003.00168.x
• Grudkowska M, Zagdanska B (2004) Multifunctional role of plant cysteine proteinases. Acta Biochim Pol 51:609–624
• Grudkowska M, Zagdanska B (2010) Acclimation to frost alters proteolytic response of wheat seedlings to drought. J Plant Physiol 167:1321–1327. doi: 10.1016/j.jplph.2010.05.019
• Hara-Nishimura I, Shimada T, Hiraiwa N, Nishimura M (1995) Vacuolar processing enzyme responsible for maturation of seed protein. J Plant Physiol 145:6412–6417. doi: 10.1016/S0176-1617(11)81275-7
• Hawkes S, Li H, Taniguchi G (2010) Zymography and reverse zymography for detecting MMPs and TIMPs. In: Clark IM (ed) Matrix metalloproteinase protocols, vol 622. Humana Press, New Jersey, pp 257–269. doi: 10.1007/978-1-60327-299-5_16
• Janska H, Kwasniak M, Szczepanowska J (2013) Protein quality control in organelles—AAA/FtsH story. Biochim Biophys Acta-Mol Cell Res 1833:381–387. doi: 10.1016/j.bbamcr.2012.03.016
• Joo AR, Jeya M, Lee KM, Sim WI, Kim JS, Kim IW, Kim YS, Oh DK, Gunasekaran P, Lee JK (2009) Purification and characterization of a beta-1,4-glucosidase from a newly isolated strain of Fomitopsis pinicola. Appl Microbiol Biot 83:285–294. doi: 10.1007/s00253-009-1861-7
• Kwon M-A, Kim HS, Hahm D-H, Song JK (2011) Synthesis activity-based zymography for detection of lipases and esterases. Biotechnol Lett 33:741–746. doi: 10.1007/s10529-010-0485-4
• Lantz MS, Ciborowski P (1994) Zymographic techniques for detection and characterization of microbial proteases. In: Clark VL, Bavoil PM (eds) Methods of enzymology, vol 235. Academic Press, New York, pp 563–594. doi: 10.1016/0076-6879(94)35171-6
• Larocca M, Rossano R, Riccio P (2010) Analysis of green kiwi fruit (Actinidia deliciosa cv. Hayward) proteinases by two-dimensional zymography and direct identification of zymographic spots by mass spectrometry. J Sci Food Agric 90:2411–2418. doi: 10.1002/jsfa.4100
• Lucinski R, Jackowski G (2013) AtFtsH heterocomplex-mediated degradation of apoproteins of the major light harvesting complex of photosystem II (LHCII) in response to stresses. J Plant Physiol 170:1082–1089. doi: 10.1016/j.jplph.2013.1003.1008
• Maciel F, Salles CC, Retamal C, Gomes V, Machado OT (2011) Identification and partial characterization of two cysteine proteases from castor bean leaves (Ricinus communis L.) activated by wounding and methyl jasmonate stress. Acta Physiol Plant 33:1867–1875. doi: 10.1007/s11738-011-0730-z
• Marek L (2013) The role of the apoptosome in the activation of procaspase-9. Postep Hig Med Dosw 67:54–64 (in Polish)
• Mariano G, Funk C (2012) Matrix metalloproteinases in plants: a brief overview. Physiol Plant 145:196–202. doi: 10.1111/j.1399-3054.2011.01544.x
• Misas-Villamil JC, Toenges G, Kolodziejek I, Sadaghiani AM, Kaschani F, Colby T, Bogyo M, Van Der Hoorn RAL (2013) Activity profiling of vacuolar processing enzymes reveals a role for VPE during oomycete infection. Plant J 73:689–700. doi: 10.1111/tpj.12062
• Ouyang L, Shi Z, Zhao S, Wang FT, Zhou TT, Liu B, Bao JK (2012) Programmed cell death pathways in cancer: a review of apoptosis, autophagy and programmed necrosis. Cell Prolif 45:487–498. doi: 10.1111/j.1365-2184.2012.00845.x
• Pan D, Hill AP, Kashou A, Wilson KA, Tan-Wilson A (2011) Electrophoretic transfer protein zymography. Anal Biochem 411:277–283. doi: 10.1016/j.ab.2011.01.015
• Paparelli E, Gonzali S, Parlanti S, Novi G, Giorgi FM, Licausi F, Kosmacz M, Feil R, Lunn JE, Brust H, van Dongen JT, Steup M, Perata P (2012) Misexpression of a chloroplast aspartyl protease leads to severe growth defects and alters carbohydrate metabolism in Arabidopsis. Plant Physiol 160:1237–1250. doi: 10.1104/pp.112.204016
• Piechota J, Kolodziejczak M, Juszczak I, Sakamoto W, Janska H (2010) Identification and characterization of high molecular weight complexes formed by matrix AAA proteases and prohibitins in mitochondria of Arabidopsis thaliana. J Biol Chem 285:12512–12521. doi: 12510.11074/jbc.M12109.063644
• Piszczek E, Dudkiewicz M, Sobczak M (2011) Molecular cloning and phylogenetic analysis of cereal type II metacaspase cDNA from wheat. Biol Plant 55:614–624. doi: 10.1007/s10535-011-0159-7
• Piszczek E, Dudkiewicz M, Mielecki M (2012) Biochemical and bioinformatic Characterization of type ii metacaspase protein (TaeMCAII) from wheat. Plant Mol Biol Rep 30:1338–1347. doi: 10.1007/s11105-012-0450-6
• Rawlings ND, Barrett AJ, Bateman A (2012) MEROPS: the database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Res 40:D343–D350 (http://merops.sanger.ac.uk). doi: 10.1093/nar/gkr987
• Richau KH, Kaschani F, Verdoes M, Pansuriya TC, Niessen S, Stüber K, Colby T, Overkleeft HS, Bogyo M, van der Hoorn RAL (2012) Subclassification and biochemical analysis of plant papain-like cysteine proteases displays subfamily-specific characteristics. Plant Physiol 158:1583–1599. doi: 10.1104/pp.112.194001
• Robinson DB, Glew RH (1980) Acid phosphatase in Gaucher’s disease. Clin Chem 26:371–382
• Rossano R, Larocca M, Riccio P (2011) 2-D zymographic analysis of broccoli (Brassica oleracea, L. var. Italica) florets proteases: follow up of cysteine protease isotypes in the course of post-harvest senescence. J Plant Physiol 168:1517–1525.
• Royer JC, Nakas JP (1990) Simple, sensitive zymogram technique for detection of xylanase activity in polyacrylamide gels. Appl Environ Microbiol 56:1516–1517
• Sakamoto K, Toyohara H (2009) A comparative study of cellulase and hemicellulase activities of brackish water clam Corbicula japonica with those of other marine Veneroida bivalves. J Exp Biol 212:2812–2818. doi: 10.1242/jeb.031567
• Shrestha A, Megeney L (2012) The non-death role of metacaspase proteases. Front Oncol 2:78. doi: 10.3389/fonc.2012.00078
• Shukla MR, Yadav R, Desai A (2009) Catalase and superoxide dismutase double staining zymogram technique for Deinococcus and Kocuria species exposed to multiple stresses. J Basic Microbiol 49:593–597. doi: 10.1002/jobm.200900037
• Srivalli B, Khanna-Chopra R (2001) Induction of new isoforms of superoxide dismutase and catalase enzymes in the flag leaf of wheat during monocarpic senescence. Biochem Biophys Res Commun 288:1037–1042. doi: 0.1006/bbrc.2001.5843
• Srivastava A, Nair J, Bendigeri D, Vijaykumar A, Ramaswamy N, D’Souza S (2009) Purification and characterization of a salinity induced alkaline protease from isolated spinach chloroplasts. Acta Physiol Plant 31:187–197. doi: 10.1007/s11738-008-0219-6
• Steup M, Gerbling K (1983) Multiple forms of amylase in leaf extracts: electrophoretic transfer of the enzyme forms into amylose-containing polyacrylamide gels. Anal Biochem 134:96–100. doi: 10.1016/0003-2697(83)90268-3
• Tamura T, Terauchi K, Kiyosaki T, Asakura T, Funaki J, Matsumoto I, Misaka T, Abe K (2007) Differential expression of wheat aspartic proteinases, WAP1 and WAP2, in germinating and maturing seeds. J Plant Physiol 164:470–477.
• Tsiatsiani L, Van Breusegem F, Gallois P, Zavialov A, Lam E, Bozhkov P (2011) Metacaspases. Cell Death Differ 18:1279–1288. doi: 10.1038/cdd.2011.66
• Vandooren J, Geurts N, Martens E, Van den Steen PE, Opdenakker G (2013) Zymography methods for visualizing hydrolytic enzymes. Nat Methods 10:211–220. doi: 10.1038/nmeth.237
• Wagner R, Aigner H, Funk C (2012) FtsH proteases located in the plant chloroplast. Physiol Plant 145:203–214. doi: 10.1111/j.1399-3054.2011.01548.x
• Wang J, Bayles KW (2013) Programmed cell death in plants: lessons from bacteria? Trends Plant Sci 18:133–139. doi: 10.1016/j.tplants.2012.09.004
• Wilkesman JG, Schroder HC (2007) Analysis of serine proteases from marine sponges by 2-D zymography. Electrophoresis 28:429–43610. doi: 1002/elps.200600332
• Zhang N, Jones BL (1995) Characterization of germinated barley endoproteolytic enzymes by two-dimensional gel electrophoresis. J Cereal Sci 21:145–153. doi: 10.1016/0733-5210(95)90030-6
• Zhang N, Jones BL (1996) Purification and partial characterization of a 31-kDa cysteine endopeptidase from germinated barley. Planta 199:565–572. doi: 10.1007/bf00195188

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