RESONANCE RAMAN-SPECTROSCOPIC STUDIES OF LIGATED MUTANT MYOGLOBINS

被引:5
作者
BIRAM, D [1 ]
HESTER, RE [1 ]
机构
[1] UNIV YORK,DEPT CHEM,YORK YO1 5DD,N YORKSHIRE,ENGLAND
来源
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY | 1994年 / 1204卷 / 02期
关键词
MYOGLOBIN; LIGAND BINDING; MUTANT; RAMAN SPECTROSCOPY;
D O I
10.1016/0167-4838(94)90010-8
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
Ligand binding (CO and N-3(-)) to wild-type porcine myoglobin and to several mutant forms, expressed and purified from E. coli cells, has been studied using Raman spectroscopy. The v(Fe-CO) stretching vibration in Mb(II)CO has been compared for the wild-type and mutant proteins. This gives a broad band consisting of five components, indicating five possible configurations of the bound CO. The distal pocket mutants show large variations in bandshape, the major component occurring at progressively lower wavenumber in the order: wild-type (WT) > E11 Val --> Thr (VT) > E7 His --> Val (HV) > the double mutant VT/HV (M2). Changes observed in the Raman band assigned to the azide bending mode in Mb(III)N(3) have been interpreted in terms of resonance structures involving two forms of azide binding. Repulsion between the bound azide ligand and the OH group of the adjacent thr residue in the VT mutant, and a shorter Fe-N(his) bond in the proximal mutant Ser --> Leu (F7), both affect this bonding. In the wild-type protein (WT), hydroxymetmyoglobin exists in a spin-state equilibrium which, at room temperature, is predominantly high-spin. In the F7 mutant this equilibrium is shifted in favour of the low-spin form. A low-spin iron species also exists in the aquometmyoglobin form of this mutant.
引用
收藏
页码:207 / 216
页数:10
相关论文
共 32 条
[21]   STEREOCHEMISTRY OF COOPERATIVE EFFECTS IN HAEMOGLOBIN [J].
PERUTZ, MF .
NATURE, 1970, 228 (5273) :726-&
[22]   NEUTRON-DIFFRACTION REVEALS OXYGEN-HISTIDINE HYDROGEN-BOND IN OXYMYOGLOBIN [J].
PHILLIPS, SEV ;
SCHOENBORN, BP .
NATURE, 1981, 292 (5818) :81-82
[23]   STRUCTURE AND REFINEMENT OF OXYMYOGLOBIN AT 1.6-A RESOLUTION [J].
PHILLIPS, SEV .
JOURNAL OF MOLECULAR BIOLOGY, 1980, 142 (04) :531-554
[24]  
ROHLFS RJ, 1990, J BIOL CHEM, V265, P3168
[25]   DISTAL POCKET POLARITY IN LIGAND-BINDING TO MYOGLOBIN - STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF A THREONINE68(E11) MUTANT [J].
SMERDON, SJ ;
DODSON, GG ;
WILKINSON, AJ ;
GIBSON, QH ;
BLACKMORE, RS ;
CARVER, TE ;
OLSON, JS .
BIOCHEMISTRY, 1991, 30 (25) :6252-6260
[26]  
SPIRO TG, 1988, BIOL APPLICATIONS RA, V3
[27]  
SPRINGER BA, 1989, J BIOL CHEM, V264, P3057
[28]   MODE OF ATTACHMENT OF AZIDE ION TO SPERM WHALE METMYOGLOBIN [J].
STRYER, L ;
WATSON, HC ;
KENDREW, JC .
JOURNAL OF MOLECULAR BIOLOGY, 1964, 8 (01) :96-&
[29]   TEMPERATURE-DEPENDENCE OF RESONANCE RAMAN-SPECTRA OF METMYOGLOBIN AND METHEMOGLOBIN AZIDE - DETECTION OF RESONANCE-ENHANCED BOUND AZIDE VIBRATIONS AND IRON-AZIDE STRETCH [J].
TSUBAKI, M ;
SRIVASTAVA, RB ;
YU, NT .
BIOCHEMISTRY, 1981, 20 (04) :946-952
[30]   RESONANCE RAMAN INVESTIGATION OF CARBON-MONOXIDE BONDING IN (CARBON MONOXY) HEMOGLOBIN AND (CARBON MONOXY) MYOGLOBIN - DETECTION OF FE-CO STRETCHING AND FE-C-O BENDING VIBRATIONS AND INFLUENCE OF THE QUATERNARY STRUCTURE CHANGE [J].
TSUBAKI, M ;
SRIVASTAVA, RB ;
YU, NT .
BIOCHEMISTRY, 1982, 21 (06) :1132-1140