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

Content Provider	Springer Nature Link
Author	Grudkowska, Małgorzata Lisik, Piotr Rybka, Krystyna
Copyright Year	2013
Abstract	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.
Starting Page	3477
Ending Page	3482
Page Count	6
File Format	PDF
ISSN	01375881
Journal	Acta Physiologiae Plantarum
Volume Number	35
Issue Number	12
e-ISSN	18611664
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2013-09-12
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	1D zymography 2D zymography Proteinases Wheat Plant Physiology Plant Genetics & Genomics Plant Biochemistry Plant Pathology Plant Anatomy/Development Agriculture
Content Type	Text
Resource Type	Article
Subject	Physiology Plant Science Agronomy and Crop Science

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