PROTEIN PROTEIN-INTERACTION OF DETERGENT-SOLUBILIZED CA-2+-ATPASE DURING ATP HYDROLYSIS ANALYZED BY LOW-ANGLE LASER-LIGHT SCATTERING PHOTOMETRY COUPLED WITH HIGH-PERFORMANCE GEL CHROMATOGRAPHY

被引:10
作者
KIJIMA, Y
TAKAGI, T
SHIGEKAWA, M
TADA, M
机构
[1] OSAKA UNIV, SCH MED,DEPT MED 1,1-1-50 FUKUSHIMA,FUKUSHIMA KU, OSAKA 553, JAPAN
[2] OSAKA UNIV, SCH MED, DEPT PATHOPHYSIOL, OSAKA 553, JAPAN
[3] OSAKA UNIV, INST PROT RES, SUITA, OSAKA 565, JAPAN
[4] NATL CARDIOVASC CTR, RES INST, DEPT MOLEC PHYSIOL, SUITA, OSAKA 565, JAPAN
关键词
ATPase; Ca[!sup]2+[!/sup]-; Dimeric structure; Laser light scattering; Protein-protein interaction; Sarcoplasmic reticulum;
D O I
10.1016/0167-4838(90)90114-U
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
Protein-protein interaction of detergent-solubilized Ca2+-ATPase was examined, employing low-angle light scattering photometry coupled with high-performance gel chromatography. When solubilized with octa(ethylene glycol) mono-n-dodecyl ether (C12E8) and chromatographed in the presence of 0.3 mg/ml C12E8, the Ca+-ATPase emerged as a single peak with an intermediate molecular weight between the monomer and the dimer, showing a dissociation-association equilibrium of the two components. In the presence of 50 μg/ml phosphatidylcholine and 0.3 mg/ml C12E8 at 0°C, the Ca2+-ATPase (0.8 mg) emerged as the two distinct components with molecular weights of 125 000±2100 (n = 3) and 211 300±7300 (n = 3), indicating that there was no rapid interconversion between the monomer and the dimer. Under the latter conditions, addition of ATP induced fusion of the two components. The apparent molecular weight of the fused peak shifted from the monomer to the dimer as the amount of protein increased. Addition of ADP or adenosine 5′-(β, γ-methylene triphosphate), however, did not induce such fusion of the peaks.The ATP-induced fusion of the peaks was not observed either in 5 mM CaCl2, the conditions in which the rate of ATP hydrolysis was extremely slow. Thus, the solubilized Ca2+-ATPase underwent a rapid interconversion between the monomer and the dimer during ATP hydrolysis. These results suggest that the protein-protein interaction during ATP hydrolysis is an intrinsic nature of Ca2+-ATPase and that such interaction may be important for Ca2+ transport by Ca2+-ATPase in the sarcoplasmic reticulum membranes. © 1990.
引用
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页码:1 / 8
页数:8
相关论文
共 34 条
[11]   MINIMUM ENZYME UNIT FOR NA+/K+-ATPASE IS THE ALPHA-BETA-PROTOMER - DETERMINATION BY LOW-ANGLE LASER-LIGHT SCATTERING PHOTOMETRY COUPLED WITH HIGH-PERFORMANCE GEL CHROMATOGRAPHY FOR SUBSTANTIALLY SIMULTANEOUS MEASUREMENT OF ATPASE ACTIVITY AND MOLECULAR-WEIGHT [J].
HAYASHI, Y ;
MIMURA, K ;
MATSUI, H ;
TAKAGI, T .
BIOCHIMICA ET BIOPHYSICA ACTA, 1989, 983 (02) :217-229
[12]   MOLECULAR-WEIGHTS OF ALPHA-BETA-PROTOMERIC AND OLIGOMERIC UNITS OF SOLUBLE (NA+,K+)-ATPASE DETERMINED BY LOW-ANGLE LASER-LIGHT SCATTERING AFTER HIGH-PERFORMANCE GEL CHROMATOGRAPHY [J].
HAYASHI, Y ;
TAKAGI, T ;
MAEZAWA, S ;
MATSUI, H .
BIOCHIMICA ET BIOPHYSICA ACTA, 1983, 748 (02) :153-167
[13]  
HAYASHI Y, 1989, METHOD ENZYMOL, V172, P514
[14]  
HYMEL L, 1984, J BIOL CHEM, V259, P4890
[15]   MECHANISM OF CALCIUM-TRANSPORT [J].
INESI, G .
ANNUAL REVIEW OF PHYSIOLOGY, 1985, 47 :573-601
[16]   ESTIMATION OF MOLECULAR-WEIGHTS OF MEMBRANE-PROTEINS IN THE PRESENCE OF SDS BY LOW-ANGLE LASER-LIGHT SCATTERING COMBINED WITH HIGH-PERFORMANCE POROUS SILICA-GEL CHROMATOGRAPHY - CONFIRMATION OF THE TRIMER STRUCTURE OF PORIN OF THE ESCHERICHIA-COLI OUTER-MEMBRANE [J].
KAMEYAMA, K ;
NAKAE, T ;
TAKAGI, T .
BIOCHIMICA ET BIOPHYSICA ACTA, 1982, 706 (01) :19-26
[17]   USE OF GEL CHROMATOGRAPHY FOR DETERMINATION OF SIZE AND MOLECULAR-WEIGHT OF PROTEINS - FURTHER CAUTION [J].
LEMAIRE, M ;
RIVAS, E ;
MOLLER, JV .
ANALYTICAL BIOCHEMISTRY, 1980, 106 (01) :12-21
[18]   RETENTION OF ENZYME-ACTIVITY BY DETERGENT-SOLUBILIZED SARCOPLASMIC CA2+-ATPASE [J].
LEMAIRE, M ;
MOLLER, JV ;
TANFORD, C .
BIOCHEMISTRY, 1976, 15 (11) :2336-2342
[19]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[20]   STATE OF AGGREGATION OF DETERGENT-SOLUBILIZED SARCOPLASMIC-RETICULUM ADENOSINE-TRIPHOSPHATASE INVESTIGATED BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY [J].
LUDI, H ;
HASSELBACH, W .
JOURNAL OF CHROMATOGRAPHY, 1984, 297 (AUG) :111-117