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International Journal of Scientific Research in Environmental Sciences (IJSRES), 1 (12), pp. 351-356, 2013 Available online at http://www.ijsrpub.

com/ijsres ISSN: 2322-4983; 2013 IJSRPUB http://dx.doi.org/10.12983/ijsres-2013-p351-356

Full Length Research Paper Studying the Relationship between Acyl-CoA-Binding Protein 2 and Lysophospholipase 2 in Arabidopsis thaliana and Their Importance in Recycling of Cadmium
Shahin Gavanji1*, Mohsen Doostmohammadi2, Amin Mojiri3, Alireza Jalali Zand4
Young Researcher Club, Khorasgan Branch, Islamic Azad University, Khorasgan, Iran Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran 3 School of Civil Engineering, Engineering Campus, Universiti Sains Malaysia, 14300 Nibong Tebal, Penang, Malaysia 4 Department of Plant Protection, Khorasgan (Isfahan) Branch, Islamic Azad University, Isfahan, Iran *Corresponding Author: Email: shahin.gavanji@yahoo.com
2 1

Received 9 September 2013; Accepted 28 October 2013

Abstract. A major environmental concern due to dispersal of industrial and urban wastes generated by human activities is the contamination of soil. A wide range of inorganic and organic compounds cause contamination, these include heavy metals, combustible and putriscible substances, hazardous wastes, explosives and petroleum products. Major component of inorganic contaminates are heavy metals .Phytoremediation is a group of technologies that used by plants to remove, degrade or immobilize environmental toxins. This technology largely focused on the use of plants for accelerating degradation of organic contaminants. But in addition to use of plant for soil remediation, hyper accumulator plants can be used for recycling of metals from soil. Hyper accumulator plant express special proteins that help them to tolerate the high concentration of metals.in this experiment we discussed the relationship between acyl-CoA-binding protein 2 and lysophospholipase 2 and showed why this two proteins work together. Key words: phytoremediation, immobilize, toxin, organic contaminants

1. INTRODUCTION Evolution of human life led to the development of technologies and appearance of secure sciences (Bennett et al., 2003). Global development, created treatable issues especially in the area of environment protection and natural resources. Since the industrial revolution, soil contamination by toxic metals has been severely increased. According to the Ryagys article 90% of metal diffusion happened by artificial sources (Ikhuoria et al., 2000). Trace metals, including those defined as heavy, arise from industrial and mining activities discharge into coastal waters and estuaries at many sites. Any metallic element that has a high density and is toxic is called heavy metal. These anthropogically derived inputs can accumulate in local sediments and invertebrates living on or in food, and the rate of accumulation caries widely between species and heavy metal concentration found in clean conditions (Harris and Santos, 2000). Soil and water contaminated with metals include a major environmental and human health problem that is still in need of an effective technological solution. Nonradioactive As, Cd, Cu, Hg, Pb and Zn and radioactive Sr, Cs and U are the most environmentally important metallic pollutants. Microbial

bioremediation has been somewhat successful for the degradation of certain organic contaminants, but is ineffective at addressing the challenge of toxic metal contamination, particularly in soil (Dudka and Miller, 1999). Today it is clear that human activities lead to increasing the metal pollutant in soil and water (Nriagu, 1996). Mining and smelting operations are important causes of heavy metal contamination in the environment due to activities such as mineral excavation, ore transportation, smelting and refining, and disposal of the tailings and waste waters around mines (Dudka and Adriano, 1997; Navarro et al., 2008). A lot number of chemical materials, heavy metals and other industrial wastes discharged in coastal regions. This toxic materials spread in environment and thought the food chain lead to toxic effects on human and animals (Dembitsky, 2003; Manohar et al., 2006). Ordinarily the soil is a place for disposal of many heavy metals. Today the methods for reducing metal pollutants in soil are expensive method and damage to environmetnt.so finding low cost methods for disposing of heavy metals (BIO-WISE, 2003 ; Aboulroos et al., 2006) Metal-binding proteins play critical catalytic, regulatory and structural roles in the cells. (Formigari et al., 2007) Their Identication and characterization can contribute toward a better use of


Gavanji et al. Studying the Relationship between Acyl-CoA-Binding Protein 2 and Lysophospholipase 2 in Arabidopsis thaliana and Their Importance in Recycling of Cadmium

them in metal pollutant remediation. ACBPs are implicated in acyl-CoA trafcking in many eukaryotes and some prokaryotes. Six genes encode proteins designated as AtACBP1AtACBP6 in the Arabidopsis thaliana ACBP family. These ACBPs are conserved in the acylCoA-binding domain. In Arabidopsis, the ACBP family consists of six members (AtACBP1AtACBP6) with AtACBP6 being the smallest (10.4 kDa) and AtACBP4 the largest (73.1 kDa). As in stress full conditions like metal pollutant environments the coding of Acyl-CoA binding protein increased we try to understand the effect of this change (Blanvillain et al., 2009). The use of plants to remove heavy metals from contaminated soils can be also applied as a tool for extracting metals, especially trace metals from soil. Today Phytostabilisation is mostly used for the remediation of soil, sediment and sledges (Mueller et al. 1999) and depends on roots

ability to limit contaminant mobility and bioavailability in the soil (Berti et al., 2000). It is a good approach to remove the contamination primarily from soil and isolate it, without destroying the soil structure and fertility. The removed heavy metal can be recycled from the contaminated plant biomass (Brooks et al., 1998). Factors such as growth rate, element selectivity, resistance to disease, method of harvesting, are also important (Cunningham et al., 1996). However slow growth, shallow root system, small biomass production, final disposal limit the use of hyper accumulator species (Brooks et al., 1994). In this study we tried to understand the mechanism of cadmium tolerance in plant Arabidopsis thaliana. By finding the mechanisms of tolerance in hyper accumulator plants and strengthen this pathways can be hoped to produce more resistant plant and overcame the problems that mentioned.

Fig. 1: Amino acid sequence of Acyl-CoA-binding domain-containing protein 2 1.1. Heavy metals Heavy metals are the main mineral contaminants and due to the use of sludge or compost from urban excrement, pesticides and fertilizers, heavy metals published in the environment and pollutions are causing significant portion of earth (Halim and Conte, 2003) Although metals are naturally present in the Earths crust and many of them are essential for cellular processes but all metals are toxic in high concentrations (Xiaoe Yang et al., 2005). Heavy metals in soil can be a danger for the health of human and animals. As, Cd, Cr, Cu, Pb, Mg, Ni, Se, Ag and Zn are the metals that have this bad effects (McIntyre, 2003).Other metals that have such effects are Al, Cs, Co, Mn and Sr. All plants can absorb and accumulate essential metals. Plants for growth need to several concentrations of any types of metals (Long, 2002). This ability of plants let them to accumulate other non-essential metals. When the concentration of metals in plants cells become more than their threshold, it inhibit the normal function of cytoplasmic enzymes and causes cell toxicity and also through transporting nutrients in place of cation exchange cause indirect toxicity (Djingova and Kuleff, 2002).

Fig. 2: 3 dimensional structure of Acyl-CoA-binding domain-containing protein 2


International Journal of Scientific Research in Environmental Sciences (IJSRES), 1 (12), pp. 351-356, 2013

2. MATERIALS AND METHODS Acyl-CoA binding protein sequence was obtained from NCBI GenBank (figure 1), Protein Data Bank (PDB) and the 3D structure of lysophosphatidylcholine obtained from chemspider website (figure 2). In order to design the 3D structure for Acyl-CoA binding protein first of all PSI-BLAST performed and the 3D structure of homologues of acyl-CoA binding protein obtained. For designing 3D structure Easy modeler software was used. This software according to 3D structure of homologues proteins released a new 3D structure for unknown protein. Phyre2, SAM-T08 and m4t servers also used

for predicting 3D structure of Acyl-CoA binding protein and the best model with the lowest Q-mean score accepted. Energy minimization performed using UCSF chimera candidate version1.5.3. During minimization step update interval was 10 and step size was 0.02. After that, this protein was docked with lysophosphatidylcholine using Molegro software. Molegro Virtual Docker is an integrated platform for predicting protein - ligand interactions. Molegro Virtual Docker handles all aspects of the docking process from preparation of the molecules to determination of the potential binding sites of the target protein, and prediction of the binding modes of the ligands (http://www.molegro.com/trial.php).

Table 1: Amino acid sequence analysis of Metal transporter Nramp1

3. RESULTS AND DISCUSSIONS Metalloproteins are a large and diverse class of proteins which bind one or more metal ions in their native con- formation (1).A metal binding site typically consists of an ion bound to one or more protein residues (called ligands). Among the 20 amino acids, the four most common ligands are cysteine (C), histidine (H), aspartic acid (D), and glutamic acid (E). Highly conserved residues are more likely to be involved in the coordination of a metal ion, although in the case of cysteines, conservation is also often associated with the presence of a disulde bridge (a covalent bond between the sulfur atoms of two cysteines). In this study we tried to understand if there is any interaction between acyl-CoA binding protein and lysophosphatidylcholine. The secondary structure of acyl-CoA binding protein constructed with Easy Modeler software and showed with its three cofactors(SO4) in figure 1.this protein docked by Molegro software with lysophosphatidylcholine and

The results of this software indicate that acyl-CoA binding protein has high affinity to lysophosphatidylcholine. The moldocke scores for 10 times Molegro running were all lower than -80 kj/mol. As this docking studies have low moldocke scores, so it is clear that there is a direct relationship between cadmium tolerance and acyl-CoA binding protein expression. 5. CONCLUSION There are many proteins including P1B-type heavy metal ATPases, ABC transporter, phytochelatins, methallothioneins and oxidative stress-related proteins have been associated with heavy metal stress (Wei Gao et al., 2010) Some Heavy metals including Cu(II), iron [Fe(III)], nickel [Ni(II)], Cd(II) and Zn(II) are known to induce oxidative stress at high concentrations(Guerrero et al., 2006). In vitro observations showed that in response to metal stress the expression levels of lysophospholipase increased.


Gavanji et al. Studying the Relationship between Acyl-CoA-Binding Protein 2 and Lysophospholipase 2 in Arabidopsis thaliana and Their Importance in Recycling of Cadmium

Also we know that Arabidopsis thaliana lysophospholipase 2 (lysoPL2) binds acyl-CoAbinding protein 2 (ACBP2) to mediate cadmium [Cd(II)] tolerance (Larsen et al., 2006).Our results show that acyl-CoA binding protein has high affinity to lysophosphatidylcholine with average energy 83Kj/mol.So we discussed that in order to decrease

the digestive effect of increase in lysophospholipase level on cell membrane, the acyl-CoA binding protein attached to lysophospholipase 2 and by saturating lysophospholipase activation site moderate its digestive effects and increased Cd tolerance in Arabidopsis thaliana.

Fig. 3: The secondary structure of acyl-CoA binding protein and lysophosphatidylcholine (yellow)

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International Journal of Scientific Research in Environmental Sciences (IJSRES), 1 (12), pp. 351-356, 2013

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Gavanji et al. Studying the Relationship between Acyl-CoA-Binding Protein 2 and Lysophospholipase 2 in Arabidopsis thaliana and Their Importance in Recycling of Cadmium

Shahin Gavanji graduated in Biotechnology at MSc at the Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran. He has over 10 international medals in invention. Shahin Gavanji's research has focused on Pharmacy and Pharmacology, Nano Biotechnology, Bioinformatics, Biotechnology - Medical Biotechnology. He is editor in chief of International Journal of Scientific Research in Inventions and New Ideas.

Mohsen Doostmohammadi graduated in Biotechnology at MSc at the Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran.

Amin Mojiri is a PhD candidate in Environmental Engineering, School of Civil Engineering, Universiti Sains Malaysia (USM), Pulau Pinang. He is fellowship holder and research assistant at the School of Civil Engineering (USM). He has more than 18 articles in the international journals. He is editor and reviewer of some international journals. His area of specialization is waste management, waste recycling, wastewater treatment, wastewater recycling, and soil pollutions.

Associate Professor of Entomology, Department of Plant Protection, Khorasgan Branch, Islamic Azad University, Isfahan, Iran