Simultaneous Separation of Taxon-Specific Crystallins from Mule Duck and Characterization of Their Enzymatic Activities and Structures
學年 105
學期 2
出版（發表）日期 2017-05-17
作品名稱 Simultaneous Separation of Taxon-Specific Crystallins from Mule Duck and Characterization of Their Enzymatic Activities and Structures
作品名稱（其他語言）
著者 Chih-Hsien Wang; Chia-Chi Huang; Wenlung Chen
單位
出版者
著錄名稱、卷期、頁數 Journal of Chromatography B 1053, p.34-41
摘要 Methods to obtain pure proteins in large amounts are indispensible in protein research. We report here a large-scale/simultaneous isolation of taxon-specific crystallins (ɛ- and δ-crystallin) from the eye lenses of Mule duck. We also investigate the compositions, enzymatic activities, and structures of these purified taxon-specific proteins. A relatively mild method of ion-exchange chromatography was developed to fractionate ɛ-crystallin and δ-crystallin in large amount, ca. ∼6.60 mg/g-lens and ∼41.0 mg/g-lens, respectively. Both crystallins were identified by electrophoresis, HPLC, and MALDI-TOF-MS. ɛ-Crystallin, with native composition of Mr 142 kDa, consisted of two subunits of 35 kDa and 36 kDa, while δ-Crystallin, with native molecular mass of 200 kDa, comprised single subunit of Mr ∼50 kDa. Both ɛ- and δ-crystallin were tetramers. The former showed lactate dehydrogenase (LDH) activity, while the latter appeared slightly active in an argininosuccinate lyase (ASL) assay. Raman spectroscopic results indicated that the secondary structures of ɛ- and δ-crystallin were predominantly α-helix as evidenced by the vibrational stretching of amide III over 1260 cm−1 and amide I at 1255 cm−1, in greatly contrast to the anti-parallel β-sheet of α- and β-crystallin as demonstrated by amide III at 1238 cm−1 and amide I at 1672 cm−1. The microenvironments of aromatic amino acids and the status of thiol groups also vary in different crystallins. The compositions, enzyme activities, and structures of the ɛ- and δ-crystalline of Mule duck are different from those of Muscovy duck (Cairina moschata) or Kaiya duck (Anas Platyrhynchos var. domestica), which reflect faithfully species specificity.
關鍵字 Taxon-specific protein;Ion-exchange chromatography;ɛ-Crystallin;δ-Crystallin;FT-Raman;Enzyme activity
語言 en_US
ISSN 1873-376X
期刊性質 國外
收錄於 SCI
產學合作
通訊作者
審稿制度 是
國別 NLD
公開徵稿
出版型式 ,電子版,紙本
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