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Subunit Characteristics of Pig Pancreas Ferritin Revealed by MALDI-TOF MS and RP-HPLC
Authors:HUANG Lin    LIN Zhi-cao  LIN Qing  LUO Lian-zhong  HUANG He-qing    Key Laboratory of the MOD for Cell Biology  Tumor Cell Engineering  School of Life Sciences  State Key Laboratory of Physical Chemistry of Solid Surface  Key Laboratory of Chemical Biology of Fujian Province  College Chemistry & Chemical Engineering  Xiamen University  Xiamen  P R China
Institution:HUANG Lin1,3,LIN Zhi-cao1,LIN Qing1,LUO Lian-zhong1,3 , HUANG He-qing1,2,3 1. Key Laboratory of the MOD for Cell Biology , Tumor Cell Engineering,School of Life Sciences,2. State Key Laboratory of Physical Chemistry of Solid Surface,3. Key Laboratory of Chemical Biology of Fujian Province,College Chemistry & Chemical Engineering,Xiamen University,Xiamen 361005,P. R. China
Abstract:Pig pancreas ferritin(PPF) was purified by ultra-centrifugation,ion-exchange chromatography,and native gradient polyacrylamide gel electrophoresis(PAGENG).Sodium dodecyl sulfate(SDS)-PAGE indicates that PPF consists of two subunit types,namely,H(21000) and L(19000) subunits,and its core shows an average element composition of 1698 Fe3 and 179 phosphate molecules within the hollow shell,giving a 9.5:1 ratio of Fe3 to phosphate.An off line approach combining reversed-phase high-performance liquid chromatography(RP-HPLC) with matrix-assisted laser desorption ionization time of flight mass speetrometry(MALDI-TOF MS) made the decomposition of PPF shell into H and L subunits for the analysis of mass spectrometry(MS),giving molecular weights of both H(21014.4) and L(18319.9)subunits.Both subunit types were further identified by an approach combining peptide mass fingerprint(PMF) with database search.A ratio of IH to 2L subunits in PPF was determined by SDS-PAGE,RP-HPLC,and MALDI-TOF MS,respectively.It is well known that the non-covalent interaction of L-L or H-L subunits is stronger than that of H-H subunits in PPF,which may be further used to explain the unclear physiological function between H and L subunits in PPF.
Keywords:Ferritin  Pig pancreas  Subunit structure  Proteomic  MALDI-TOF MS  RP-HPLC
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