Protein Extraction and Identification by Gel Electrophoresis and Mass Spectrometry from Edible bird’s Nest Samples
Tóm tắt
Edible bird’s nest (EBN) is a delicacy rich in protein and carbohydrate from the salivary secretion of swiftlets. There are limited studies on the protein profile of EBN, mainly due to its complexity in chemical composition and diversity of species, as well as the capacity of analytical techniques. The protein extraction methods, namely, ultrasonic and detergent (Triton X-100 and SDS)-assisted methods, as well as Tris–HCl buffer solubilization were used to compare the protein profiles of EBNs harvested from two locations (Batu Pahat and Kota Tinggi) in Malaysia. Ultrasonic assisted extraction produced the highest protein content because EBN contains mostly water-soluble protein. The electrophoretic gels revealed that EBNs from Batu Pahat (17–150 kDa) exhibited more protein bands than those samples from Kota Tinggi (25–154 kDa). The difference could be explained by the variance in the geographical origin of EBNs. Additional protein bands (25, 27, and 92 kDa) were detected for the detergent-assisted methods. These could be poorly water-soluble membrane proteins that were released after cell lysis by detergents. The mass spectra revealed that acidic mammalian chitinase precursor is the most abundant protein. The other proteins include trypsinogen, inter-alpha-trypsin inhibitor heavy chain H2 precursor, tumor necrosis factor receptor superfamily member 5 precursor, and phospholipase A2-like. There are also collagen and co-enzyme Q-binding proteins which are important for skin complexion in line with the traditional belief of Chinese community. The protein extraction methods could produce good quality of proteins for affirmative confirmation using gel electrophoresis and mass spectrometric identification.
Tài liệu tham khảo
Bahuguna R, Purohit MC, Nautiyal BP, Prakash V, Nautiyal MC, Purohit AN (2003) Polypeptide patterns and isoenzymes in different populations of Aconitum heterophyllum wall ex Royle, from Garhwal Himalaya. Physiol Mol Biol Plants 9:95–100
Berezovski MV, Mak TW, Krylov SN (2007) Cell lysis inside the capillary facilitated by transverse diffusion of laminar flow profiles (TDLFP). Anal Bioanal Chem 387:91–96
Bollag DM, Rozycki MD, Edelstein SJ (1996) Protein methods: second edition. Wiley. Publication, New York
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Brown RB, Audet J (2008) Current techniques for single-cell lysis. J Roy Soc Interface 5:S131–S138
Chan KM, Zhang Z (2012) Leucine-rich repeat and WD repeat-containing protein 1 is recruited to pericentric heterochromatin by trimethylated lysine 9 of histone H3 and maintains heterochromatin silencing. J Biol Chem 287:15024–15033
Chua KH, Lee TH, Kamini N, Nor HMY, Lee CT, Eddie TTT, Ramlan AA (2013) Edible Bird’s Nest as a chondro-protective agent for human chondrocytes isolated from osteoarthritic knee: in vitro study. BMC Complement Altern Med 13:19
Chua LS, Nurulaini AR, Bustanur R, Lee CT (2012) Plant proteins, minerals and trace elements of Eurycoma longifilia (Tongkat ali). Nat Prod Res 27(4–5):314–318
Contreras L, Ritter A, Dennett G, Boehmwald F, Guitton N, Pineau C, Moenne A, Potin P, Correa JA (2008) Two dimensional gel electrophoresis analysis of brown algal protein extracts. J Phycol 44:1315–1321
Dennis EA, Cao J, Hsu YH, Magrioti V, Kokotos G (2011) Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Chem Rev 111:6130–6185
Fraser JR, Laurent TC, Laurent UB (1997) Hyaluronan: its nature, distribution, functions and turnover. J Intern Med 242(1):27–33
Friedenauer S, Berlet HH (1989) Sensitivity and variability of the Bradford protein assay in the presence of detergents. Anal Biochem 78:263–268
Guo CT, Takahashi T, Bukawa W, Takahashi N, Yagi H, Kato K, Miyamoto D, Suzuki D, Suzuki Y (2006) Edible bird’s nest extract inhibits influenza virus infection. Antivir Res 70:140–146
Heringdorf DM, Jakobs KH (2007) Lysophospholipid receptors: signalling, pharmacology and regulation by lysophospholipid metabolism. Biochim Biophys Acta 1768:923–940
Herrera-Estrella A, Chet I (1999) Chintinases in biological control. In: Jolles P, Muzzarelli RAA (eds) Chitin and chitinase. Birkhauser, Basel, pp. 171–184
Kang N, Hails CJ, Sigurdsson JB (1991) Nest construction and egg-laying in edible-nest swiflets Aerodramus spp. and the implications for harvesting. Ibis 133(2):170–177
Kathan RH, Weeks DI (1969) Structure studies of collocalia mucoid: I. Carbohydrate and amino acid composition. Arch Biochem Biophys 134:572–576
Kechrid Z, Bouzerna N (2004) Effect of zinc deficiency on zinc and carbohydrate metabolism in genetically diabetic C57BL/Ksj Db+/Db+) and non-diabetic original strain (C57BL/Ksj) mice. Turk J Med Sci 34:367–373
Kong YC, Keung WM, Yip TT, Ko KM, Tsao SW, Ng MH (1987) Evidence that epidermal growth factor is present in swiftlet’s (Collocalia) nest. Comp Biochem Physiol Biochem Mol Biol 87:221–226
Koon LC, Cranbrook E (2002) Swiftlets of Borneo—builders of edible nests. Natural History Publication (Borneo) SDN. BHD., Sabah, pp. 1–171
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Liu X, Lai X, Zhang S, Huang X, Lan Q, Li Y, Li BC, Zhang Q, Hong D, Yang G (2012) Proteomic profile of edible bird’s nest. J Agric Food Chem 60:12477–12481
Ma F, Liu D (2012) Sketch of the edible bird’s nest and its important bioactivities. Food Res Int 48:559–567
Marcone MF (2005) Characterization of the edible bird’s nest the “Caviar of the East”. Food Res Int 38:1125–1134
Marni S, Marzura MR, Norzela AM, Khairunnisak M, Bing CH, Eddy AA (2014) Preliminary study on free sialic acid content of edible bird nest from Johor and Kelantan. Malaysian J Vet Res 5:9–14
Matsukawa N, Matsumoto M, Bukawa W, Chiji H, Nakayama K, Hara H, Tsukahara T (2011) Improvement of bone strength and dermal thickness due to dietary edible bird’s nest extract in ovariectomized rats. Biosci Biotechnol Biochem 75:590–592
Murphy RM, Lamb GD (2013) Important considerations for protein analyses using antibody based techniques: down-sizing Western blotting up-sizes outcomes. J Physiol 591(Pt 23):5823–5831
Nakagawa H, Hama Y, Sumi T, Li SC, Maskos K, Kalayanamitra K, Mizumoto S, Sugahara K, Li YT (2007) Occurrence of a non-sulfated chondroitin proteoglycan in the dried saliva of Collocalia swiftlets (edible bird’s-nest). Glycobiology 17:157–164
Qiu R, Yang Y, Zhao H, Li J, Xin Q, Shan S, Liu Y, Dang J, Yu X, Gong Y, Liu Q (2013) Signal transducer and activator of transcription 6 directly regulates human ORMDL3 expression. FEBS J 280:2014–2026
Rajapakse S, Ogiwara K, Takahashi T (2014) Characterization and expression of trypsinogen and trypsin in medaka testis. Zool Sci 31(12):840–848
Roh KB, Lee J, Kim YS, Park J, Kim JH, Lee J, Park D (2012) Mechanisms of edible bird’s nest extract-induced proliferation of human adipose-derived stem cells. Evid Based Complement Alt Med 2012:Article ID 797520, 11 pages.
Sazili AQ, Noor Azihan MZ, Shuhaimi M, Hilmi M, Panandam JM (2010) The effectiveness of soluble protein extractability under the effect of pH, molarity, type of buffers of three different major skeletal muscles in cattle. Asian J Biol Sci 1–5
Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins from silver stained polyacrylamide gels. Anal Chem 68:850–858
Sims RW (1961) The identification of Malaysian species of swiftlets. Ibis 103a(2):205–210
Utomo B, Rosyidi D, Radiati LE, Puspaningsih NNT, Proborini WD (2014) Protein characterization of extracted water from three kinds of edible bird nest using SDS-PAGE CBB staining and SDS-PAGE glycoprotein staining and LC-MS/MS analyses. IOSR J Agr Vet Sci 7:33–38
Vimala B, Hussain H, Wan Nazaimoon WM (2011) Effects of edible bird’s nest on tumour necrosis factor-alpha secretion, nitric oxide production and cell viability of lipopolysaccharide-stimulated RAW 264.7 macrophages. Food Agric Immunol 23:303–314
Yan H, Lamm ME, Bjorling E, Huang YT (2002) Multiple functions of immunoglobulin A in mucosal defense against viruses: an in vitro measles virus model. J Virol 76(21):10972–10979
Yew MY, Koh RY, Chye SM, Othman I, Ng KY (2014) Edible bird’s nest ameliorates oxidative stress-induced apoptosis in SH-SY5Y human neuroblastoma cells. BMC Complement Altern Med 2014(14):391