Heterogeneity and pigment composition of isolated photosystem II reaction centers

被引:25
作者
Zheleva, D [1 ]
Hankamer, B [1 ]
Barber, J [1 ]
机构
[1] UNIV LONDON IMPERIAL COLL SCI TECHNOL & MED,DEPT BIOCHEM,WOLFSON LABS,LONDON SW7 2AY,ENGLAND
关键词
D O I
10.1021/bi961382h
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Photosystem II reaction centers (RC) isolated from peas (Pisum sativum L) purified by ion-exchange chromatography were shown, by high-performance liquid chromatography (HPLC) size-exclusion analyses, to consist of a mixture of monomers (180 +/- 20 kDa) and dimers (390 +/- 35 kDa). Both fractions were resolved and purified by sucrose density gradient centrifugation and their homogeneity was demonstrated in size-exclusion HPLC elution profiles, Also present in the nonresolved preparation and the monomeric fraction were low levels of CP47 apoprotein (1.8% and 0.9% apoprotein of that found in a CP47-RC preparation). This CP47 contamination could maximally account for 0.41 and 0.22 Chl/RC, respectively, based on 22 chlorophylls being bound to each CP47 protein. The level of CP47 apoprotein was undetectable in the dimeric fractions. Pigment analysis using reverse-phase HPLC confirmed that contamination by chlorophyll bound to the CP47 apoprotein in the nonresolved RC preparation was low and that the ratio of chlorophyll a to pheophytin a remained 6 when the preparation was separated into its monomeric and dimeric components. We conclude, in agreement with earlier work, that the reaction center of PSII, when isolated using mild detergents and ion-exchange chromatography, contains 6 chlorophyll a/2 pheophytin a. We therefore do not concur with the recent published work of Pueyo et al. [(1995) Biochemistry 34, 15214-15218) that this type of preparation contains 4 chlorophyll a/2 pheophytin a and that the remaining 2 chlorophyll a are due to contamination by CP47.
引用
收藏
页码:15074 / 15079
页数:6
相关论文
共 30 条
[1]   DISINTEGRATION AND RECONSTITUTION OF PHOTOSYSTEM-II REACTION CENTER CORE COMPLEX .1. PREPARATION AND CHARACTERIZATION OF 3 DIFFERENT TYPES OF SUBCOMPLEX [J].
AKABORI, K ;
TSUKAMOTO, H ;
TSUKIHARA, J ;
NAGATSUKA, T ;
MOTOKAWA, O ;
TOYOSHIMA, Y .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 932 (03) :345-357
[2]   CORE ANTENNA COMPLEXES, CP43 AND CP47, OF HIGHER-PLANT PHOTOSYSTEM-II - SPECTRAL PROPERTIES, PIGMENT STOICHIOMETRY, AND AMINO-ACID-COMPOSITION [J].
ALFONSO, M ;
MONTOYA, G ;
CASES, R ;
RODRIGUEZ, R ;
PICOREL, R .
BIOCHEMISTRY, 1994, 33 (34) :10494-10500
[3]   ISOLATION AND CHARACTERIZATION OF THE PHOTOSYSTEM 2 REACTION-CENTER COMPLEX FROM A DOUBLE MUTANT OF CHLAMYDOMONAS-REINHARDTII [J].
ALIZADEH, S ;
NIXON, PJ ;
TELFER, A ;
BARBER, J .
PHOTOSYNTHESIS RESEARCH, 1995, 43 (02) :165-171
[4]   PIGMENT STOICHIOMETRY OF THE PHOTOSYSTEM-II REACTION-CENTER FROM HIGHER-PLANTS [J].
AURED, M ;
MOLINER, E ;
ALFONSO, M ;
YRUELA, I ;
TOON, SP ;
SEIBERT, M ;
PICOREL, R .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 1994, 1187 (02) :187-190
[5]   CHARACTERIZATION OF A PS-II REACTION CENTER ISOLATED FROM THE CHLOROPLASTS OF PISUM-SATIVUM [J].
BARBER, J ;
CHAPMAN, DJ ;
TELFER, A .
FEBS LETTERS, 1987, 220 (01) :67-73
[6]   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
[7]   EXCITON LEVEL STRUCTURE AND DYNAMICS IN THE CP47 ANTENNA COMPLEX OF PHOTOSYSTEM-II [J].
CHANG, HC ;
JANKOWIAK, R ;
YOCUM, CF ;
PICOREL, R ;
ALFONSO, M ;
SEIBERT, M ;
SMALL, GJ .
JOURNAL OF PHYSICAL CHEMISTRY, 1994, 98 (31) :7717-7724
[8]   ELECTRON-TRANSPORT PROPERTIES OF THE ISOLATED D1-D2-CYTOCHROME-B-559 PHOTOSYSTEM-II REACTION CENTER [J].
CHAPMAN, DJ ;
GOUNARIS, K ;
BARBER, J .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 933 (03) :423-431
[9]  
CHAPMAN DJ, 1989, PHOTOSYNTHETICA, V23, P411
[10]  
CHAPMAN DJ, 1991, METHODS PLANT BIOCH, V5, P171