Subunit Characteristics of Pig Pancreas Ferritin Revealed by MALDI-TOF MS and RP-HPLC

IF 3.1 4区化学 Q2 CHEMISTRY, MULTIDISCIPLINARY

Chemical Research in Chinese Universities Pub Date : 2008-09-01 DOI:10.1016/S1005-9040(08)60116-6

Lin HUANG , Zhi-cao LIN , Qing LIN , Lian-zhong LUO , He-qing HUANG

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引用次数: 6

Abstract

Pig pancreas ferritin(PPF) was purified by ultra-centrifugation, ion-exchange chromatography, and native gradient polyacrylamide gel electrophoresis(PAGE_NG). 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 spectrometry(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 1H 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.

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利用MALDI-TOF质谱和反相高效液相色谱分析猪胰腺铁蛋白亚基特征

采用超离心、离子交换色谱、天然梯度聚丙烯酰胺凝胶电泳等方法纯化猪胰腺铁蛋白(PPF)。十二烷基硫酸钠(SDS)-PAGE表明PPF由H(21000)和L(19000)两个亚基类型组成，其核心显示空心壳内平均元素组成为1698个Fe3+和179个磷酸盐分子，Fe3+与磷酸盐的比例为9.5:1。采用反相高效液相色谱(RP-HPLC)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)相结合的离线方法，将PPF壳分解为H和L亚基进行质谱分析，得到H(21014.4)和L(18319.9)亚基的分子量。采用肽质量指纹图谱(PMF)和数据库检索相结合的方法进一步鉴定了两种亚基类型。采用SDS-PAGE、RP-HPLC和MALDI-TOF MS分别测定PPF中1H与2L亚基的比值。众所周知，PPF中L-L或H-L亚基的非共价相互作用强于H-H亚基，这可能进一步解释了PPF中H和L亚基之间不明确的生理功能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Chemical Research in Chinese Universities 化学-化学综合

CiteScore

5.30

自引率

6.50%

发文量

152

审稿时长

3.0 months

期刊介绍： The journal publishes research articles, letters/communications and reviews written by faculty members, researchers and postgraduates in universities, colleges and research institutes all over China and overseas. It reports the latest and most creative results of important fundamental research in all aspects of chemistry and of developments with significant consequences across subdisciplines. Main research areas include (but are not limited to): Organic chemistry (synthesis, characterization, and application); Inorganic chemistry (bio-inorganic chemistry, inorganic material chemistry); Analytical chemistry (especially chemometrics and the application of instrumental analysis and spectroscopy); Physical chemistry (mechanisms, catalysis, thermodynamics and dynamics); Polymer chemistry and polymer physics (mechanisms, material, catalysis, thermodynamics and dynamics); Quantum chemistry (quantum mechanical theory, quantum partition function, quantum statistical mechanics); Biochemistry; Biochemical engineering; Medicinal chemistry; Nanoscience (nanochemistry, nanomaterials).