GST profile expression study in some selected plants:in silico approach

Soma Banerjee • Riddhi Goswami

Received: 9 March 2009 / Accepted: 15 September 2009 / Published online: 15 October 2009

� Springer Science+Business Media, LLC. 2009

Abstract Glutathione acts as a protein disulfide reductant,

which detoxifies herbicides by conjugation, either sponta-

neously or by the activity of one of a number of glutathione-

S-transferases (GSTs), and regulates gene expression in

response to environmental stress and pathogen attack. GSTs

play role in both normal cellular metabolism as well as in

the detoxification of a wide variety of xenobiotic com-

pounds, and they have been intensively studied with regard

to herbicide detoxification in plants. A newly discovered

plant GST subclass has been implicated in numerous stress

responses, including those arising from pathogen attack,

oxidative stress, and heavy-metal toxicity. In addition, plant

GSTs play a role in the cellular response to auxins and

during the normal metabolism of plant secondary products

like anthocyanins and cinnamic acid. The present study

involves two in silico analytical approaches—general sec-

ondary structure prediction studies of the proteins and

detailed signature pattern studies of some selected GST

classes in Arabdiopsis thaliana, Mustard, Maize, and Bread

wheat by standard Bioinformatics tools; structure prediction

tools; signature pattern tools; and the evolutionary trends

were analyzed by ClustalW. For this purpose, sequences

were obtained from standard databases. The study reveals

that these proteins are mainly alpha helical in nature with

specific signature pattern similar to phosphokinase C,

tyrosine kinase, and casein kinase II proteins, which are

closely related to plant oxidative stress. This study aims to

comprehend the relationship of GST gene family and plant

oxidative stress with respect to certain specific conserved

motifs, which may help in future studies for screening of

biomodulators involved in plant stress metabolism.

Keywords GST family � Structure prediction �Evolutionary trend � Signature pattern � Conserved motif

Introduction

The increased production of toxic oxygen derivatives is

considered to be a universal or common feature of stress

conditions. Plant and other organisms have evolved a wide

range of mechanisms to contend with this problem. The

antioxidant system in plants with its wide array of anti-

oxidant enzymes plays a pivotal role in these defence

mechanisms. The effects of the action of free radicals on

membranes include the induction of lipid peroxidation and

fatty acid de-esterification. Both ethylene biosynthesis and

membrane breakdown, which appear to be closely linked,

seem to involve free radicals, although the sequence of

events generating these free radicals is still poorly under-

stood. It is clear that the capacity and activity of the anti-

oxidative defence system are important in limiting

oxidative damage and in destroying active oxygen species

that are produced in excess of those normally required for

metabolism.

Glutathione-S-transferase (GST) belongs to a family of

isoenzymes [1] that catalyze the glutathione conjugation of

a variety of electrophilic compounds, including carcino-

gens, mutagens, cytotoxic drugs, and their metabolites, and

detoxification products of reactive oxidation. Their pres-

ence in plants was first recognized shortly afterwards in

1970, when a GST activity from maize was shown to be

responsible for conjugating the chloro-S-triazine atrazine

with reduced glutathione (GSH), thereby protecting the

S. Banerjee (&) � R. Goswami

Department of Biotechnology, Heritage Institute of Technology,

Chowbaga Road, Anandapur, P.O. East Kolkata Township,

Kolkata 700107, India

e-mail: somabanerjee2005@gmail.com

123

Mol Cell Biochem (2013) 380:283–300

DOI 10.1007/s11010-009-0258-3

crop from injury by this herbicide [2]. Since that time, GST

activities, or the corresponding enzymes or gene sequen-

ces, have been identified in all animals, plants, and fungi

analyzed to date [3, 4]. In addition to the dimeric soluble

GSTs, other proteins have been identified as having a

restricted ability to conjugate xenobiotics (foreign organic

compounds) with GSH, notably the distantly related

mitochondrial kappa GSTs and the trimeric microsomal

GSTs of animals [5, 6].

GSTs are dimeric enzymes that show five independent

classes, alpha, mu, pi, sigma [7–10], and theta [7, 8, 10].

Classes alpha, mu, and pi initially proposed by Mannervick

et al. in 1988 [7], are present only in animals and yeasts but

absent in bacteria and plants. The very heterogeneous theta

class, reported by Meyer et al. 1991 [7, 11, 12] are present in

yeasts, plants, bacteria, rats, humans, chickens, salmon, and

non vertebrates such as flies and apparently absent in lower

animals such as mollusks, nematodes, and platyhelminthes

[13]. Zeta and theta GSTs are found in both animals and

plants, but the tau and phi classes are plant-specific.

Each soluble GST is a dimer of approximately 26 kDa

sub-units, typically forming a hydrophobic 50 kDa protein

with an isoelectric point in the pH range 4–5. In the case of

phi and tau GSTs, only subunits from the same class will

dimerize [14, 15]. Within a class, however, the subunits

can dimerize even if they are quite different in amino acid

sequence [15]. As determined for the GSTs active in her-

bicide metabolism in maize and wheat, the ability to form

heterodimers greatly increases the diversity of the GSTs in

plants [16], but the functional significance of this mixing

and matching of subunits has yet to be determined.

Structural information on plant GSTs is available for phi

GSTs from Arabidopsis thaliana [17] and maize [18, 19]

and for a zeta-class GST from Arabidopsis thaliana [20].

Despite the extreme sequence divergence between the GST

classes the overall structures of the enzymes are remark-

ably similar. Some of the structural characteristics of GSTs

are also observed in other GSH-dependent proteins, such as

glutaredoxin, suggesting a strong evolutionary pressure to

retain structural motifs involved in binding GSH at the

active site [4, 6, 21].

Materials and methods

Nucleotide sequences for different GST subclasses were

retrieved from the NCBI nucleotide databank National

Center for Biotechnology Information (http://www.ncbi.

nlm.nih.gov/) (Table 1).

The plants chosen for this study are: Arabidopsis thali-

ana, Oryza sativa, Triticum aestivum, and Brassica juncea.

The retrieved data were then submitted to GENSCAN to

get corresponding protein sequences and their secondary

structures were obtained from the various secondary

structure prediction tools using three different online soft-

wares; viz., GOR IV, HNN, and SOPMA (http://www.ex

pasy.org/tools/). All these softwares are algorithm based

and have their own set of strengths and weaknesses. Thus,

an average of results obtained from all these three soft-

wares was taken into consideration.

The results obtained with these programs were taken for

further analysis of the primary folding pattern. The curated data

in its FASTA format was submitted to the Scan prosite soft-

ware (http://www.expasy.org/tools/scanprosite/) for searching

out the fingerprints. From the Scan prosite results the con-

served domains having minimum length of eight amino acids

were considered for further analysis.

Furthermore the phylogenetic analysis would suggest

that all soluble GSTs have arisen from an ancient pro-

genitor evaluated with the help of the online software

ClustalW (http://align.genome.jp/).

Results

Secondary structure analysis

Table 2 shows the secondary structure prediction (in %) of

the proteins belonging to TAU, PHI, ZETA, and THETA

classes in all the selected plant species as obtained from the

above-mentioned softwares. It shows that for Tau class, ahelix ranges from 40.42 to 51.99% in Arabidopsis thaliana,

48.39 to 54.82% in Oryza sativa, 38.92 to 52.75% in

Triticum aestivum, and Random coils ranges from 27.97 to

37.71% in Arabidopsis thaliana, 27.65 to 38.94% in Oryza

sativa, 28.29 to 48.55% in Triticum aestivum where in the

whole group, a helix is predominant. It shows that for Phi

class, a helix ranges from 40.56 to 42.36% in Arabidopsis

thaliana, 42.20 to 46.20% in Oryza sativa, 42.14 to 46.78%

in Triticum aestivum and 45.69 to 47.59% in Brassica

juncea. Random coils ranges from 33.17 to 42.69% in

Arabidopsis thaliana, 35.24 to 43.69% in Oryza sativa,

33.99 to 44.10% in Triticum aestivum, 32.58 to 43.04% in

Brassica juncea where in the whole group, a helix is pre-

dominant It shows that for Theta class, a helix ranges from

39.77 to 51.43% in Arabidopsis thaliana, 36.98 to 43.46%

in Oryza sativa Random coils ranges from 35.80 to 47.75%

in Arabidopsis thaliana, 35.24 to 43.69% in Oryza sativa,

in the whole group, a helix is predominant It shows that for

Zeta class, a helix ranges from 37.88 to 48.30% in

Arabidopsis thaliana, 39.41 to 47.33% in Oryza sativa

where in the whole group, a helix is predominant.

From the percentage of occurrence of the above three

secondary structures it was observed that all the three classes

of proteins have a tendency to have alpha (a) helical struc-

ture followed by random coil and then by beta (b) sheet.

284 Mol Cell Biochem (2013) 380:283–300

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Table 1 Nucleotide sequence information (Retrieved from NCBI)

Organism Class Accession number/gi no. Class Accession number/gi no. Class Accession number/gi no.

Arabidiopsis thaliana TAU gi|145362079| PHI gi|145360494| THETA gi|145358713|

gi|145360447| gi|145339930| gi|145358712|

gi|145360446| gi|145336565| gi|30693768|

gi|186504025| gi|145329994|

gi|186504020| gi|79316324|

gi|145338288| gi|42567907|

gi|145337707| gi|30690771|

gi|145337706| gi|30684734|

gi|145337353| gi|30678782|

gi|145337352| gi|30678436|

gi|145336873| gi|30678435|

gi|145335813| gi|30678431|

gi|145335812| gi|30678427|

gi|42569442| gi|30678032|

gi|42565447| gi|18412490|

gi|42561897| gi|18379032|

gi|30699325|

gi|30699029| ZETA gi|186498763|

gi|30697687| gi|145359839|

gi|30696330| gi|145358829

gi|30689787| gi|42570652|

gi|30689782| gi|18395358|

gi|30685181| gi|28932691|

gi|30684248| gi|11967658|

gi|30684233|

gi|30681676|

gi|18411918|

gi|18411911|

gi|18404581|

Oryza sativa TAU gi|46404432I PHI gi|46276328I THETA gi|46404430I

gi|46276332I gi|46276326I gi|15430708I

gi|46276330I gi|11177844I

gi|33304611I gi|11177842I ZETA gi|46243592I

gi|33304609I gi|11177840I gi|85700989I

gi|11177828I gi|15430706I

gi|11177832I gi|1177838I

gi|1177834I

gi|11177836I

Triticum aestivum TAU gi|20067422| PHI gi|23572871|

gi|20067420| gi|23504748|

gi|20067418| gi|23504746|

gi|20067416| gi|23504744|

gi|20067414| gi|23504742|

gi|23504740|

gi|23504738|

Mol Cell Biochem (2013) 380:283–300 285

123

After detailed analysis of the Scan prosite result of

different classes of GST in all the chosen plant species it

was found that the sequences belonged to the following

families (Tables 3, 4) CK2_PHOSPHO_SITE (Casein

Kinase II Phosphorylation site), TYR_PHOSPHO_SITE

(Tyrosine Kinase Phosphorylation site), Myristyl

(n-myristoylation site), and PKC_PHOSPHO_SITE (protein

kinase c phosphorylation site). Only the families

CK2_PHOSPHO_SITE (Casein Kinase II Phosphorylation

site), TYR_PHOSPHO_SITE (Tyrosine Kinase Phosphor-

ylation site), and PKC_PHOSPHO_SITE (protein kinase c

phosphorylation site) are chosen because of their direct

relationship with GSTs role in cellular regulation and

metabolism. These conserved sequences are of 3, 4, and 7

amino acid lengths and are present in all the selected plant

species. Casein Kinase II is known to have possible

mechanism of gene activation of salicylic acid in tobacco

[21]. This conserved sequence is of four amino acid length

and is present in most of the selected plant species. This

sequence is found to be variable at its 2nd, 3rd positions

but depending upon the class this variation also changes

from one class of protein to another. For example for both

Tau and Phi class the signature pattern is more or less

conserved, but for Theta and Zeta there is more variation.

This sequence is found to be variable at its 2nd and 3rd

positions. This variation also changes from one class of

protein to another. In plant protein kinase C (PKC) has

been found to be involved in many aspects of cellular

regulation and metabolism [22]. This conserved sequence

is of three amino acid lengths and is present in most of the

selected plant species. This sequence is found to be vari-

able at its 2nd positions but also depends upon the class.

Like for both Theta and Phi class the signature pattern is

more or less conserved with the only exception in

Table 1 continued

Organism Class Accession number/gi no. Class Accession number/gi no. Class Accession number/gi no.

Brassica jun�cea PHI gi|31790104|

gi|31790102|

gi|31790098|

gi|31790096|

gi|31790094|

gi|31790092|

Table 2 Comparative analysis of secondary structures of GST classes

Class Organism GOR IV SOPMA HNN

a-helix (%) b-sheet (%) Random

coils (%)

a-helix (%) b-sheet (%) Random

coils (%)

a-helix

(%)

b-sheet

(%)

Random

coils (%)

TAU Arabidopsis thaliana 40.42 16.16 27.97 51.99 12.32 27.97 47.75 15.21 37.71

Oryza sativa 48.39 12.68 38.94 54.82 11.05 27.65 49.91 11.67 38.43

Triticum aestivum 38.92 12.54 48.55 52.75 12.82 28.29 47.46 10.69 41.85

FINAL 42.58 12.61 43.64 53.19 12.06 27.97 48.37 12.52 39.33

PHI Arabidopsis thaliana 40.56 16.75 42.69 40.46 16.21 33.17 42.36 13.79 42.29

Oryza sativa 46.20 12.46 41.35 42.20 14.16 35.24 45.55 10.76 43.69

Triticum aestivum 42.14 13.77 44.10 45.54 13.93 33.99 46.78 9.94 43.28

Brassica juncea 45.69 11.27 43.04 46.59 14.08 32.58 47.59 10.25 42.16

FINAL 43.65 13.56 42.80 43.70 14.60 33.75 45.57 11.19 42.86

THETA Arabidopsis thaliana 46.12 11.80 42.07 51.43 11.67 31.47 39.77 13.01 47.23

Oryza sativa 36.98 14.81 48.22 43.46 17.42 31.50 43.05 15.58 41.37

FINAL 41.55 13.31 45.15 47.45 14.55 32.40 41.41 14.30 44.30

ZETA Arabidopsis thaliana 37.88 14.38 47.75 48.30 10.31 35.80 41.69 15.22 43.09

Oryza sativa 39.41 15.10 45.50 47.33 13.21 33.70 39.41 15.10 45.50

FINAL 38.65 14.74 46.63 47.82 11.76 34.75 40.55 15.16 44.30

286 Mol Cell Biochem (2013) 380:283–300

123

Table 3 Fingerprint analysis

showing signature position of

different GST proteins in

different organisms

gi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

A. PHI protein

Arabdiopsis thaliana

gi|145360494| 19–22 12–14

88–91 13–15

151–154 199–201

162–165 208–210

gi|145339930| 131–134 15–17

167–170 185–187 154–160

190–192

203–205

208–210

gi|145336565| 166–169

182–185 153–159

236–239

gi|145329994| 19–22 12–14

88–91

gi|79316324| 181–184 31–33

49–51

78–80

96–98

199–201

gi|42567907| 163–166 153–155

178–181 200–202

204–206

gi|30690771| 21–24 3–5

64–67 15–17

165–168 80–82

201–203

gi|30684734| 19–22 12–14

88–91 199–201

162–165

gi|30678782| 163–166 124–126

187–190 153–155 206–214

187–189

200–202

gi|30678436| 183–186 33–35

51–53

80–82

98–100

201–203

gi|30678435| 181–184 31–33

49–51

78–80

96–98

199–201

Mol Cell Biochem (2013) 380:283–300 287

123

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

gi|30678431| 89–92 15–17

131–134 80–82 150–156

163–166 89–91

128–130

181–183

199–201

gi|30678427| 132–135 15–17

164–167 80–82 151–157

129–131

160–162

182–184

200–202

204–206

gi|30678032| 167–170 14–16

15–17 154–160

185–187

190–192

203–205

208–210

gi|18412490| 26–29

63–66

86–89

101–104

164–167

gi|18379032| 200–203 22–24

50–52 35–42

126–128

132–134

156–158

218–220

227–229

Oryza sativa

giI46276328I 65–68 37–39

166–169 58–60

184–186

192–194

193–195

205–207

209–211

giI46276326I 13–16 5–7

59–62 42–44 208–214

66–69 157–159

194–197 158–160

221–224 194–196

258–261 240–242

258–260

288 Mol Cell Biochem (2013) 380:283–300

123

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

giI11177844I 38–41 186–188

65–68 193–195

169–172 206–208

210–212

giI11177842I 167–170 203–205

giI11177840I 37–40 138–140

91–94

165–168

Triticum aestivum

gi|23572871| 102–105

133–136

154–157

165–168

gi|23504748| 162–165

182–185

gi|23504746| 102–105

133–136

154–157

165–168

gi|23504744| 88–90

175–177

195–197

gi|23504742| 21–24 152–154

88–91 202–204

162–165 218–220

gi|23504740| 65–68

163–166 174–181

gi|23504738| 66–69

164–167 175–182

184–187

Brassica juncea

gi|31790104| – 161–163

– 170–172

– 175–177

– 188–190

– 193–195

gi|31790102| – 170–172

– 175–177 139–145

– 188–190

– 193–195

gi|31790098| – 15–17

– 185–187 154–160

– 190–192

– 203–205

– 208–210

Mol Cell Biochem (2013) 380:283–300 289

123

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

gi|31790096| – 170–172

– 175–177 139–145

– 188–190

– 193–195

gi|31790094| – 15–17

– 185–187 154–160

– 203–205

– 208–210

gi|31790092| – 15–17

B. THETA protein

Arabdiopsis thaliana

gi|145358713| 80–83 91–93

98–101 204–206

145–148 264–266

166–169 288–290

249–252 390–392

274–277 407–409

288–291 408–410

358–361 448–450

424–427 575–577

461–464

471–474

507–510

518–521

538–541

558–561

575–578

gi|145358712| 81–84 92–94

99–102 194–196

146–149 205–207

167–170

gi|30693768| 80–83 91–93

98–101 155–157

145–148 203–205

166–169 204–206

274–277 264–266

288–291 288–290

360–363 409–411

461–464 410–412

472–475 449–451

507–510 495–497

539–542 576–578

559–562 581–583

576–579

Oryza sativa

giI46404430I 131–134 115–117

giI15430708I 168–171 187–189

187–190

290 Mol Cell Biochem (2013) 380:283–300

123

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

Lycopersicon esculentum

gi|10567811| 117–120 116–118

199–202

gi|10567809| 16–19 76–78

127–130 92–94

165–168

gi|10567807| 3–5

39–41

78–80

gi|10567805| 115–118 15–17

194–197 119–121

176–178

194–196

gi|10567803| 37–39

219–221

C. TAU protein

Arabdiopsis thaliana

gi|145362079| 120–123 20–22

137–140

149–152

151–154

163–166

gi|145360447| 119–122 57–59

119–121

197–199

gi|145360446| 142–145 7–9

179–182 19–21

183–186 142–144

gi|186504025| 65–68 18–20

gi|186504020| 65–68 18–20

79–81

118–120

179–181

gi|145338288| 44–47 58–60

100–103 100–102

158–161

gi|145337707| 182–185 22–24

163–165

186–188

gi|145337706| 166–169 38–40

208–211 202–204

gi|145337353| 7–10

147–150

178–181

gi|145337352| 70–73 9–11

177–180 97–99

215–218 150–152

Mol Cell Biochem (2013) 380:283–300 291

123

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

gi|145336873| 42–45 205–207

120–123

gi|145335813| 117–120 111–113

187–189

203–205

gi|145335812| 162–165 109–111

198–201 202–204

gi|42569442| 68–71 97–99

120–122

gi|42565447| 2–5 85–87

156–159 119–121

179–182 184–186

203–206

gi|42561897| 40–43 111–113

gi|30699325| 116–119 79–81

146–149

198–201

gi|30699029| 127–130 19–21

84–86

gi|30697687| 94–97 20–22

111–114

123–126

125–128

137–140

gi|30696330| 42–45

gi|30689787| 175–178 6–8

213–216 18–20

238–241 29–31

240–243 99–101

131–133

148–150

gi|30689782| 67–70 6–8

174–177 18–20

212–215 29–31

98–100

147–149

gi|30685181| 77–80 109–111

201–203

209–211

gi|30684248| 142–145 19–21

142–144

gi|30684233| 5–8 21–23

82–85 29–31

121–124 43–45

82–84

121–123

gi|30681676| 40–43

gi|18411918| 119–122 205–207

292 Mol Cell Biochem (2013) 380:283–300

123

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

gi|18411911| 116–119 79–81

162–165 123–125

198–201 167–169

185–187

202–204

gi|18404581| 80–83 97–99

203–206 136–138

207–209

Oryza sativa

giI46404432I 142–145 111–113

giI46276332I 100–103 175–177

173–176

giI46276330I 6–9

125–128

giI33304611I 122–125 27–29

213–215

giI33304609I 4–7 119–121

20–23

124–127

133–136

185–188

giI11177836I 104–107 95–97

101–103

129–131

Triticum aestivum

gi|20067422| 44–47 124–126

125–128 198–200

152–155

209–212

gi|20067420| 122–125 97–99

119–121

gi|20067418| 177–180 44–46

gi|20067416| 177–180 44–46

gi|20067414| 177–180 44–46

D. ZETA protein

Arabdiopsis thaliana

gi|186498763| 4–7 30–36

145–148

160–163

gi|145359839| 198–201

4–7 30–36

152–155

167–170

gi|42570652| 205–208

50–53 33–39

148–151 31–33

163–166

Mol Cell Biochem (2013) 380:283–300 293

123

Lycopersicon esculentum, but for Tau and Zeta there has

been more variation. Tyrosine kinase signature is seven

amino acid in length and only found in Zeta class of GST.

Both in Arabidopsis thaliana and Oryza sativa this par-

ticular signature pattern is fully conserved and no variation

is observed which supports the view presented by Dixon

(2002) [23].

Phylogenetic analysis

Phylogenetic analysis revealed evolutionary relationships

for different classes of GST in plants (Fig. 1). For Phi

class, Triticum aestivum and Brassica junceaa are evolu-

tionary closely related; again Arabidopsis thaliana for tau

class and Lycopersicon esculentum in theta class are evo-

lutionary correlated; while tau class of Oryza sativa and

Triticum aestivum are closely related and share a common

evolutionary path with the Arabdiopsis thaliana for tau

class and Lycopersicon esculentum in theta class. For both

in theta and zeta class of Arabidopsis thaliana and Oryza

sativa share a common evolutionary relationship among

themselves.

Discussion

This study was designed to have a clear-cut idea about the

structural and functional correlation of different plant GST

classes in relation to plant metabolism. As evidenced from

the results of the three programs studied, it is apparent that

all the three classes of proteins have a tendency to have

alpha (a) helical structure followed by random coil and

then by beta (b) sheet.

Our aim was to search for probable colinearity between

GST classes and role of these classes in plant detoxifica-

tion, endogenous metabolism, and look for any conserved

motifs that are expressed in different plant species. The

fingerprinting analysis was done with Scan prosite, which

consists of a large collection of biologically meaningful

signatures that are described as patterns or profiles. Each

signature is linked to a documentation that provides useful

biological information on the protein family, domain or

functional site identified by the signature. Our results show

that within those sequences three major classes of signa-

tures are predominant—Casein Kinase II, PKC, and

Tyrosine kinase, which are functionally related to various

plant signaling pathways.

Table 3 continuedgi no. Position

Signature name CK2_PHOSPHO_SITE PKC_PHOSPHO_SITE TYR_PHOSPHO_SITE

gi|18395358| 201–204

4–7 30–36

145–148

160–163

gi|28932691| 198–201

4–7 30–36

145–148

160–163

gi|11967658| 198–201

Oryza sativa

giI46243592I 25–27 35–41

11–14 11–13

22–25 33–35

giI46243592I 107–110 119–121

giI85700989I 178–181 237–239

226–229

110–113

70–73

90–93

giI15430706I 4–7 30–36

giI1177838I 145–148

160–163

294 Mol Cell Biochem (2013) 380:283–300

123

In plants, Casein Kinase II appears to be involved in a

number or processes including circadian clock regulation

in Arabidopsis thaliana [24], photoperiod sensitivity in rice

[25]. Rohilla et al. [26] showed experimentally the pres-

ence of two distinct alpha subunits and two distinct beta

subunits in the purified protein complex of rice. This result

is similar to our present results which shows that the pro-

tein is mainly alpha helical in nature which in Tau is

48.05% for Phi is 44.31% for Theta is 43.47% for Zeta is

42.34%. In plants, protein phosphorylation has been

implicated in responses to many signals, including light,

pathogen invasion, hormones, temperature stress, and

nutrient deprivation. Activities of several plant metabolic

and regulatory enzymes are also controlled by reversible

phosphorylation of PKCs [22]. The present study also

revealed a probable correlation with PKC class of signa-

tures. Result of the present study revealed that on the basis

of sequence similarity and gene organization, they appear

to have evolved from a common ancestral GST into four

distinct classes, namely the Phi, Tau, Zeta, and Theta GSTs

[27], which is also similar to our phylogenetic analyses

(Fig. 1). The two largest classes are the plant-specific Phi

and Tau GSTs. Both classes have major roles in herbicide

detoxification [28]. In addition, these GSTs have less well

characterized roles in endogenous metabolism including

functioning as glutathione peroxidases counteracting oxi-

dative stress [28, 29], stress signaling proteins [30], and

regulators of apoptosis [31]. In contrast, the smaller Zeta

and Theta classes of GSTs are also found in animals and

fungi, indicating conserved and essential functions for

these enzymes in all eukaryotes. Similar result has been

observed in the phylogenetic analysis where Theta and

Zeta class of sequences are sharing common node of origin.

Moreover, only in Zeta class in both Arabdiopsis thaliana

and Oryza sativa signature pattern has been found to be

highly conserved for the Tyro_phospho_site. This finding

is in concordance with the report which shows Zeta GSTs

in Arabidopsis thaliana, animals, and fungi catalyze the

glutathione-dependent isomerization of maleylacetoacetate

to fumarylacetoacetate, an essential step in the catabolism

Table 4 Fingerprint analysis result for different GST classes

Fingerprint analysis

Signature Class Plant species Sequence

TYRO_PHOSPHO_SITE Zeta Arabidopsis thaliana K G L D Y E Y

Oryza sativa K G L E Y E Y

CK2_PHOSPHO_SITE Theta Arabidopsis thaliana [T/S] [R/I/V/E] [L/A/K] [D/E]

Oryza sativa [S] K T E

Lycopersicon esculentum [T/S] L V [D/E]

Phi Arabidopsis thaliana [T/S]L [V/A] E

Oryza sativa [T/S] L [A][D/E]

Triticum aestivum [T/S] L [V/A] [D/E]

Tau Arabidopsis thaliana [T/S][V/K/L][S/G/E/P] [D/E]

Oryza sativa [T/S] [E/S/G/L] [E/T/A][D/E]

Triticum aestivum [T/S] [L/E] [P/A] [D/E]

Zeta Arabidopsis thaliana [T/S] [L/D] A [D/E]

Oryza sativa [T/S] [E/S/N/L/K] [K/S/F/V/D] [D/E]

PKC_PHOSPHO_SITE Zeta Arabidopsis thaliana T l K

Oryza sativa [T/S][E/H/T/K] K

Theta Arabidopsis thaliana [T/S][S/K][K/R]

Oryza sativa S S R

Lycopersicon esculentum [T/S] [N/E/W][K/R]

Phi Arabidopsis thaliana [T/S][K/S/R][K/R]

Oryza sativa [T/S][K/F/S][K/R]

Triticum aestivum S A K

Brassica juncea [T/S] [K/Q] [K/R]

Tau Arabidopsis thaliana [T/S] [E/A/K/R] [K/R]

Oryza sativa [T/S] [P/F/G/W/K][K/R]

Triticum aestivum [T/S] D [K/R]

Mol Cell Biochem (2013) 380:283–300 295

123

of tyrosine [31], whereas Theta class GSTs act as potent

glutathione peroxidases detoxifying organic hydroperox-

ides formed during oxidative stress [27].

Thus, our results show the structural and functional

colinearity among the different GST classes with biologi-

cally significant signature patterns (Casein Kinase II, PKC,

Multiple sequence alignment results:

Scores Table

Fig. 1 Multiple sequence alignment results: scores table

296 Mol Cell Biochem (2013) 380:283–300

123

Alignment

Fig. 1 continued

Mol Cell Biochem (2013) 380:283–300 297

123

Fig. 1 continued

298 Mol Cell Biochem (2013) 380:283–300

123

and Tyrosine kinase) within the GST sequences in plants.

This study may help in future studies for screening of

biomodulators involved in plant stress metabolism.

Acknowledgments The authors wish to acknowledge the authori-

ties of Heritage Institute of Technology for providing necessary

technical support for this work.

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