DOI : 10.35124/bca.2020.20.1.1397 Biochem. Cell. Arch. Vol. 20, No. 1, pp. 1397-1400, 2020 www.connectjournals.com/bca ISSN 0972-5075 STUDY ON CHEMICAL COMPOSITION AND DESIGNING WEB PATTERNS OF IRAQI SPIDER SILK

Zeina N. Al-Azawii Department of Biology, Collage of Education for Pure Science (Ibn Al-Haitham), University of , Baghdad, . e-mail : [email protected] (Received 30 August 2019, Revised 30 November 2019, Accepted 7 December 2019)

ABSTRACT : Present work deals with HPLC method for determination of amino acid and study chemical composition of spider silk, this specimens were collected from variant regions in Baghdad all spiders produce silk and most produce multiple types of silk fibers. Silks are typically semicrystal line fibers that among the strongest, toughest and stretchiest biological fibers spider’s silk is made up of protein is termed a spidroin, for spider fibroin. Spidroins consist of 100–400 amino acids major amino acids in the silk proteins are alanine 24.3% and glycine 30.2% Serine and proline are also present in significant quantities in some types of silk. Glycine is rich regions give spider silk elasticity forming a morphous areas in structure, Alanine is rich regions link together form crystall ineare as that give spider silk its strength the exact composition of these proteins depends on factors including species and diet, the remaining components are mostly glutamine, serine, leucine, valine, proline, tyrosine and arginine the function of silk is prey capture webs provides a crucial link between the molecular biology of silk and spider ecology. Web building ability generally the spiders are considered as weavers or non-weavers, the weavers make the snares to trap insects for food in family Pholcidae, while the non-weavers hunt the prey by chasing in families Lycosidae, Gnaphosidae, Salticidae and Oxyopidae in this research, we study the chemical composition of spiders silk and the designing web patterns are Orb web, Horizontal webs and Single line webs, Sheet web, Irregular webs, wooly webs, Funnel webs, the orb web and sheet web so in this research, we need to know more about the spider silk and the spider webs. Key words : Alanine, glycine, spidroins, sticky silk, web patterns.

INTRODUCTION web (Blackledge et al, 2011). Dragline silk is the strongest Spider silk is incredibly tough and is stronger by kind of silk because it must support the weight of the weight than steel. Quantitatively, spider silk is five times spider (Dicko et al, 2004). Parachuting or ballooning which stronger than steel of the same diameter. It is finer than is used to aid the dispersal of young and to find new areas the human hair (most threads are a few microns in as a food source. Silk is released and is caught by the diameter) and is able to keep its strength below -40°C. wind to lift the spider up into the air - flying spiders, Spider silk is also very elastic and capture silk (sticky 5.Shelters such as burrows or nests and 6. Egg-sacs with silk for catching prey) remains unbroken after being Mating: male spiders weave sperm webs on which they stretched 2-4 times its original length. Spider silk is deposit sperm and subsequently transfer it to their front tougher, more elastic and more waterproof than silkworm palps, ready for placing on a females genital organs silk so it could have a much wider range of applications (Foelix, 1996; Dyakonov et al, 2012). Some species make (Casem et al, 1999; Dicko et al, 2004; Andersson et al, a web and coat it with sex pheromones to attract a mate. 2016). It is simple to see why spider silk is of such interest There are a few types of spider webs found in the wild, to materials chemists since new ultra-strong fibres based and many spiders are classified by the webs they weave on the silk could be developed. Spiders use silk for a different types and different in the shapes and sizesof variety of functions:1. Swathing silk for the wrapping spider webs including : Orb web associated primarily with and immobilization of prey, 2. Webs for catching prey the family Araneidae, as well as Tetragnathidae and usingsticky silk - it is elastic to prevent the prey from Uloboridae, Horizontal webs and Single line webs, Sheet rebounding off the web, 3. Draglines which are used to web, Irregular webs inthe family Theridiidae, wooly webs, connect the spider to the web, as safety lines in case a Funnel webs divided into primitive and modernin the family spider should fall and as the non-sticky spokes of the Agelenidae (Griffiths and Salanitri, 1980; Garb et al, 2010). 1398 Zeina N. Al-Azawii MATERIALS AND METHODS assessing the strengths of silk scientists usually refer to Specimen collection dragline the data we do have applies to the toughest spider silk we know of, just how much variation there is between The specimens were collected from variant region different silks we get a lot of facts about the strength of in Baghdad Al-Kadhimya, Hurriya City, Al-, spider silk that sometimes seem contradictory, although Iskan, Washash, Al-Adel, Yarmouk, Baya’, Al-Sa’adoon they all agree that Hu et al (2006), Huemmerich et al Zayouna Al-Jadriya Al-Za’franiya during 10-1- (2008) spider silk is way stronger than any other silk or 2018 to 29-12-2018. The specimens were found in leafs, natural material silks, like many other biomaterials, have bricks, wood, flagstone, trees etc. specimens were put in a hierarchical structure, the primary structure is the amino plastic box and transferred to laboratory for study. The acid sequence of its proteins (spidroin), amino acid first step is isolation the individual then but in plastic glass analysis. Multiple analyses were carried out by HPLC to were kindly supplied by water and fed diet of German Pico-Tag amino acid analysis system mainly consisting cockroaches. of highly repetitive glycine and alanine blocks, which is Silk collection and solubilization why silks are often referred to as a block co-polymer Controlled silking was performed and silking was (White et al, 1986; Pravin et al, 2010). On a secondary restricted to the spiders which were large enough to be structure level, the short side chained alanine is mainly easily manipulated without damaging the spider. The found in the crystalline domains (beta sheets) of the silking procedure averaged 30 minutes and 5.0 milligrams nanofibril, glycine is mostly found in the so-called the mature females were continuously observed the amorphous matrix consisting of helical and beta turn glandular source of silk all samples were examined using structures. Various compounds other than protein are light microscope and to preparation of silk samples for foundto enhance the fibre’s properties. Pyrrolidine has analysis on the Pico-Tag involves the following procedures hygroscopic properties which keeps the silk moist while which is: also warding off an invasion (Table 1). It occurs in (1) Conventional hydrolysis, usually in acid, to releasethe especially high concentration in glue threads. Potassium constituent amino-acids (a general procedure is hydrogen phosphate releases hydrogen ions in aqueous described). solution, resulting in a pH of about 3, making the silk acidic and protecting it from fungi and bacteria and there is (2) Preparation of the hydrolysis for chromatographyi.e. different type of amino acid in spider silk like glutamine, drying, re-drying and derivatization. serine, leucine, valine, proline, tyrosine and arginine (3) Separation of the PTC derivatives by HPLC. Silk (Lizuka, 1970; Komatzu, 1979; John et al, 2000) and there sample solubilization in (1.0-2.0 mg) of water were are seven types of silk produced by seven silk glands. A placed in pyrextes tubes are supplied with phenolic single spider does not possess all seven glands, but has caps volume 9 mlto test (White et al, 1986; Pravin et at least three if it is male (dragline, attachment and al, 2010; John et al, 2000). swathing silk) or four, if it is female. The additional one Spider webs patterns identification and study spider is for egg sac silk the seven types of gland are: achniform activities gland: swathing silk, cylindriform gland: egg sac silk ampullate glands (major and minor): non-sticky dragline The different web patterns are identified and different silk. Silk from the minor ampullate gland is only half as activities of spiders are studied which includes foraging, strong as that from the major gland, pyriform gland: egg laying and ecology of the spiders, simultaneously attaching threads - attachment discs are made, which different web patterns (Griffiths and Salanitri, 1980; Opell anchor a thread to a surface or another thread, flagelli et al, 2006). form gland: core fibres of sticky silk and aggregate gland: Photography outer part of sticky silk - droplets of an adhesive substance Pictures and comparing and analyzing them with the are deposited along the threads (Saravanan, 2006; Opell of previous work has been done on the same and different et al, 2006; Perry et al, 2010) in the author side we study web patterns of spiders are prepared to know each web the different web patterns and different activities of pattern, photograph were taken by using super–macro spiders and identified (Work and Young, 1987; Wise, lens of Canon. 1993). RESULTS AND DISCUSSION Orb webs (Fig. 1) the typical orb-weaver spiders in Spiders can produce different kinds of silks not all family Araneidaeare classic wheel or circle shaped spiders produce all the possible types of silk when networks characteristics feature of center portion four Chemical composition and designing web patterns of Iraqi spider silk 1399

Fig. 1 : Orb webs. Fig. 2 : Horizontal webs.

Fig. 3 : Single line webs. Fig. 4 : Sheet webs or tangled webs.

Fig. 5 : Irregular webs/ Space web. Fig. 6 : Woolly webs. recorded in family Theridiidae, Uloboridae. Sheet webs or Tangled webs (Fig. 4) are also known as cob webs they appear messy and shapeless web is flat with main lines running down the center these webs are found on family Theridiidae are famous for building three- dimensional space and family Linyphiidae. Irregular webs/ Space web (Fig. 5) the irregular webs, here threads are extending in all direction are built by the family Pholcidae and Family Filistatidae, family Fig. 7 : Funnel webs. Lyniphidae shows the irregular type of web. different families were recorded in this type of web Woolly Webs (Fig. 6) are distinctive not by shape Family- Aranidae wheel-shaped, family Nephilidae Large as much as by texture. These webs consist of an adhesive golden, family Tetragnathidae long-jawed and family silk that snags prey not with a sticky glue, but by Uloboridae, Cribellate or hackled electrostatically-charged silk nanofibers. Horizontalwebs or single line webs (Figss. 2, 3) Funnel webs (Fig. 7) are built in the grassland areas there is a single horizontal line, attached at both ends to sometimes a main feature of a web design and can be branches that stretchopen spaces in the forest, were quite impressive the spider uses a funnel webs for a large 1400 Zeina N. Al-Azawii Table 1 : Amino acid composition and other protein fibers (mole%) Casem M L, Turner D and Houchin K (1999) Protein and amino acid of spider silk composition of silks from the cob weaver, Latrodectus hesperus (black widow). Int. J. Biol. Macromol. 24(2-3), 103-108. Amino acid Wool keratins Spider silk Serine Fibroin Glysine 6.1 30.2 10.6 38.3 Dicko C, Vollrath F and Kenney J M (2004) Spider silk protein refolding is controlled by changing pH. Biomacromol. 5(3), 704-710. Alanine 4,4 24.3 4.9 26.4 Valine 4.6 2.4 2.5 2.4 Dyakonov T, Yang C H, Bush D, Gosangari S, Majuru S and Fatmi A (2012) Design and characterization of a silk-fibroin-based drug Leusine 6.8 4.1 1.0 1.5 delivery platform using naproxen as a model drug. J. Drug Isoleusine 2.3 1.1 1.2 1.0 Delivery, https://doi.org/10.1155/2012/490514 Serine 10.8 3.6 28.8 10.9 Foelix R (1996) Biology of spiders. New York: Oxford University Theronine 7.9 2.5 8.8 1.3 Press, Ed. I. Lysine 1.7 0.4 2.3 1.1 Garb J E, Ayoub N A and Hayashi C Y (2010) Untangling spider silk Histidine 1.2 1.1 1.1 1.2 evolution with spidroin terminaldomains. BMC Evol. Biol. 10, Arginine 6.2 6.8 2.6 1.1 243. Proline 5.9 3.6 1.5 1.1 Griffiths J R and Salanitri V R (1980) The strength of spider silk. J. Tryptophan 1.1 1.8 1.1 1.1 Mater. Sci. 15 (2), 491. Cystine 6.9 1.1 1.0 1.1 Hu X, Vasanthavada K, Kohler K, McNary S, Moore A M and Vierra Aspartic acid 4.9 2.5 17.6 1.2 C A (2006) Molecular mechanisms of spider silk. Cell Mol. Life Sci. 63(17), 1986-1999. Glutamic acid 10.1 8.6 3.5 2.0 Phenylamine 1.8 1.1 1.2 1.2 Huemmerich D, Helsen C W, Quedzuweit S, Oschmann J, Rudolph R and Scheibel T (2008) Primary structure elements of spider Tyrosine 2.5 0.1 3.2 4.5 dragline silks and their contribution to protein solubility. Biochem. number of purposes to trap insects, hideaway from prey 43(42), 13604-13612. or predators, store eggs; and in the case of some males John W, Henderson R D, Ricker B A and Cliff W (2000) Rapid, to cohabit with a female spider and wait for mating Accurate, Sensitive and Reproducible HPLC Analysis of Amino Acids Agilent Technologies, USA. time.this webs built by family Agelenidae and Lyniphidae. Komatzu K (1979) Chemist and structure of silk. J. Annul Res. Quart. CONCLUSION 13, 64-72. Spider silks represent natural composite materials that Lizuka E (1970) Conformation of silk proteins in solution and the are biopolymers and the different silk types contain at fiber-forming property of silk fibroin under shearing stress. least 2-3 distinct structural proteins, commonly referred Collagen Symposium V, III, 29-50. to as fibroins so in this study the spider silk are same Opell B D, Bond J E and Warner D A (2006) The effects of capture spiral composition and orb-web orientation on prey interception. chemical composition of amino acid and portion but the Zool. 109, 339–345. concentration are different from spider and from the Perry D J, Bittencourt D, Siltberg-Liberles J, Rech E L and Lewis R V different region and this reason are may be back to (2010) Piriform spider silk sequences reveal unique repetitive different food, different type of glands, different ecology elements. Biomacromol. 11, 3000–3006. factors and stage age of spider. Not all spiders weave Pravin B, Madhava P, Rao B M and Someswarrao N (2010) webs, spiders do not stick to their own web because only Determination of amino acid without derivatization by using the central spiral part of the web is sticky, not the HPLC - HILIC column. J. Chem. Pharm. Res. 2(2), 372-380. spokes. The spider knows where to tread webs lose their Saravanan D (2006) Spider silk structure properties and spinning. J. stickiness after about a day due to factors such as dust Text. Apparel, Tech. and Manag. 5(1), 67-73. accumulation and exposure to air. In order to save energy White J A, Hart R J and Fry J C (1986) An evaluation of the waters the spider eats its own web before making a new one so Pico-Tag system for the amino-acid analysis of food materials. J. Automatic Chem. Clin. Lab. Automation 8(4), 170-177. the protein used for the silk threads is recycled. Wise D A (1993) Spiders in ecological webs. Cambridge, UK: REFERENCES Cambridge University Press. Andersson M, Johansson J and Rising A (2016) Silk spinning in Work R W and Young C T (1987) The amino compositions of major silkworms and spiders. Int. J. Mol. Sci. 17 (8), 1290. and minor ampullate silks of certain orb-web-building spiders Blackledge T A, Kuntner M and Agnarsson I (2011) The form and (Araneae, Araneidae). J. Arachnol. 15, 65-80. function of spider orb webs: evolution from silk to ecosystems. Adv. Insect Physiol. 41, 175–262.