Abstract
Association of gp10 and gp11 (gp=gene product) is the first step in the assembly pathway of the wedge part of the baseplate of bacteriophage T4. The gp10-gp11 complex constitutes the six tail pins at the corners of the baseplate hexagon on the distal side. The stoichiometry of the subunits, gp10 and gp11, of this complex was determined in combination with sedimentation equilibrium, Edman degradation of the complex and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). From the results of Edman degradation and SDS-PAGE, the molar ratio of gp10 and gp11 was approximately 1. On the other hand, the molecular weight of the purified gp10-gp11 complex was determined by sedimentation equilibrium to be 284 000±7000, which is in good agreement with the expected value of 269 840 if the stoichiometry is 3:3. Furthermore, comparison of the results in the presence and in the absence of reducing reagent, 2-mercaptoethanol (2-ME), in SDS-PAGE revealed that two molecules of gp10 in the complex formed a disulfide bond, while the third gp10 molecule does not participate in the disulfide bond formation. Copyright (C) 2000 Elsevier Science B.V.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 286-292 |
| Number of pages | 7 |
| Journal | Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology |
| Volume | 1479 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Jun 15 2000 |
| Externally published | Yes |
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Keywords
- Analytical ultracentrifugation
- Assembly
- Bacteriophage
- Disulfide bond
- Limited proteolysis
- Stoichiometry
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
- Structural Biology
Cite this
Stoichiometry and inter-subunit interaction of the wedge initiation complex, gp10-gp11, of bacteriophage T4. / Zhao, Li; Takeda, Shigeki; Leiman, Petr; Arisaka, Fumio.
In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, Vol. 1479, No. 1-2, 15.06.2000, p. 286-292.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Stoichiometry and inter-subunit interaction of the wedge initiation complex, gp10-gp11, of bacteriophage T4
AU - Zhao, Li
AU - Takeda, Shigeki
AU - Leiman, Petr
AU - Arisaka, Fumio
PY - 2000/6/15
Y1 - 2000/6/15
N2 - Association of gp10 and gp11 (gp=gene product) is the first step in the assembly pathway of the wedge part of the baseplate of bacteriophage T4. The gp10-gp11 complex constitutes the six tail pins at the corners of the baseplate hexagon on the distal side. The stoichiometry of the subunits, gp10 and gp11, of this complex was determined in combination with sedimentation equilibrium, Edman degradation of the complex and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). From the results of Edman degradation and SDS-PAGE, the molar ratio of gp10 and gp11 was approximately 1. On the other hand, the molecular weight of the purified gp10-gp11 complex was determined by sedimentation equilibrium to be 284 000±7000, which is in good agreement with the expected value of 269 840 if the stoichiometry is 3:3. Furthermore, comparison of the results in the presence and in the absence of reducing reagent, 2-mercaptoethanol (2-ME), in SDS-PAGE revealed that two molecules of gp10 in the complex formed a disulfide bond, while the third gp10 molecule does not participate in the disulfide bond formation. Copyright (C) 2000 Elsevier Science B.V.
AB - Association of gp10 and gp11 (gp=gene product) is the first step in the assembly pathway of the wedge part of the baseplate of bacteriophage T4. The gp10-gp11 complex constitutes the six tail pins at the corners of the baseplate hexagon on the distal side. The stoichiometry of the subunits, gp10 and gp11, of this complex was determined in combination with sedimentation equilibrium, Edman degradation of the complex and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). From the results of Edman degradation and SDS-PAGE, the molar ratio of gp10 and gp11 was approximately 1. On the other hand, the molecular weight of the purified gp10-gp11 complex was determined by sedimentation equilibrium to be 284 000±7000, which is in good agreement with the expected value of 269 840 if the stoichiometry is 3:3. Furthermore, comparison of the results in the presence and in the absence of reducing reagent, 2-mercaptoethanol (2-ME), in SDS-PAGE revealed that two molecules of gp10 in the complex formed a disulfide bond, while the third gp10 molecule does not participate in the disulfide bond formation. Copyright (C) 2000 Elsevier Science B.V.
KW - Analytical ultracentrifugation
KW - Assembly
KW - Bacteriophage
KW - Disulfide bond
KW - Limited proteolysis
KW - Stoichiometry
UR - http://www.scopus.com/inward/record.url?scp=0034659760&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034659760&partnerID=8YFLogxK
U2 - 10.1016/S0167-4838(00)00015-7
DO - 10.1016/S0167-4838(00)00015-7
M3 - Article
C2 - 11004546
AN - SCOPUS:0034659760
VL - 1479
SP - 286
EP - 292
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
SN - 1570-9639
IS - 1-2
ER -