Isolation and characterization of chloroplast Photosystem II antenna of spinach by reversed-phase liquid chromatography

被引:17
作者
Zolla, L
Timperio, AM
Testi, MG
Bianchetti, M
Bassi, R
Manera, F
Corradini, D
机构
[1] Univ Tuscia, Dipartimento Sci Ambientali, I-01100 Viterbo, Italy
[2] CNR, Ist Cromatog, I-00016 Monterotondo Stn, Rome, Italy
[3] Univ Verona, Fac Sci, Dipartimento Sci Tecnol, I-37134 Verona, Italy
[4] Univ Rome La Sapienza, Dipartimento Sci Biochim A Rossi Fanelli, I-00185 Rome, Italy
关键词
antenna system; chlorophyll-proteins; HPLC; LHC II; Photosystem II; spinach;
D O I
10.1023/A:1006397419473
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
The protein components of the Photosystem II antenna system, isolated from spinach thylakoids, have been resolved by reversed-phase high performance liquid chromatography (RP-HPLC) using a butyl-silica stationary phase packed either into analytical or semi-preparative columns. Peak identification has been accomplished by a combination of various SDS-PAGE systems employing either Comassie (or silver) staining or immunological detection using polyclonal antibodies raised against LHC II and against CP29, CP26 and CP24 proteins and by aminoacid microsequence. Moreover, peak identification is consistent with the molecular masses determined by Electrospray Ionization Mass Spectrometry (HPLC-ESI-MS). The developed RP-HPLC method allows the resolution of all the protein components of the Photosystem II major Light Harvesting Complex (LHC II) and minor PS II antenna complex (CP24, CP26 and CP29) from grana membranes (BBY) and estimation of their relative stoichiometry in natural and stressed conditions, avoiding the expensive and time consuming separation procedure by sucrose-gradient ultracentrifugation and isoelectrofocusing.
引用
收藏
页码:281 / 290
页数:10
相关论文
共 38 条
[1]   CHLOROPLAST PROTEIN-PHOSPHORYLATION COUPLES PLASTOQUINONE REDOX STATE TO DISTRIBUTION OF EXCITATION-ENERGY BETWEEN PHOTOSYSTEMS [J].
ALLEN, JF ;
BENNETT, J ;
STEINBACK, KE ;
ARNTZEN, CJ .
NATURE, 1981, 291 (5810) :25-29
[2]   PROTEIN-PHOSPHORYLATION IN REGULATION OF PHOTOSYNTHESIS [J].
ALLEN, JF .
BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1098 (03) :275-335
[3]   THE DYNAMIC PHOTOSYNTHETIC MEMBRANE AND REGULATION OF SOLAR-ENERGY CONVERSION [J].
ANDERSON, JM ;
ANDERSSON, B .
TRENDS IN BIOCHEMICAL SCIENCES, 1988, 13 (09) :351-355
[4]   CHLOROPHYLL BINDING-PROTEINS WITH ANTENNA FUNCTION IN HIGHER-PLANTS AND GREEN-ALGAE [J].
BASSI, R ;
RIGONI, F ;
GIACOMETTI, GM .
PHOTOCHEMISTRY AND PHOTOBIOLOGY, 1990, 52 (06) :1187-1206
[5]   CAROTENOID-BINDING PROTEINS OF PHOTOSYSTEM-II [J].
BASSI, R ;
PINEAU, B ;
DAINESE, P ;
MARQUARDT, J .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1993, 212 (02) :297-303
[6]   A SUPRAMOLECULAR LIGHT-HARVESTING COMPLEX FROM CHLOROPLAST PHOTOSYSTEM-II MEMBRANES [J].
BASSI, R ;
DAINESE, P .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 204 (01) :317-326
[7]   BIOCHEMICAL AND FUNCTIONAL-PROPERTIES OF PHOTOSYSTEM-II IN AGRANAL MEMBRANES FROM MAIZE MESOPHYLL AND BUNDLE-SHEATH CHLOROPLASTS [J].
BASSI, R ;
MARQUARDT, J ;
LAVERGNE, J .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1995, 233 (03) :709-719
[8]   A HIGHLY RESOLVED, OXYGEN-EVOLVING PHOTOSYSTEM-II PREPARATION FROM SPINACH THYLAKOID MEMBRANES - ELECTRON-PARAMAGNETIC-RES AND ELECTRON-TRANSPORT PROPERTIES [J].
BERTHOLD, DA ;
BABCOCK, GT ;
YOCUM, CF .
FEBS LETTERS, 1981, 134 (02) :231-234
[9]   XANTHOPHYLL CYCLE COMPONENTS AND CAPACITY FOR NONRADIATIVE ENERGY-DISSIPATION IN SUN AND SHADE LEAVES OF LIGUSTRUM-OVALIFOLIUM EXPOSED TO CONDITIONS LIMITING PHOTOSYNTHESIS [J].
BRUGNOLI, E ;
CONA, A ;
LAUTERI, M .
PHOTOSYNTHESIS RESEARCH, 1994, 41 (03) :451-463
[10]  
BURNETTE WN, 1981, ANAL BIOCHEM, V112, P195, DOI 10.1016/0003-2697(81)90281-5