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Volumn 277, Issue 5325, 1997, Pages 567-570

Dual role of phosphatidylinositol-3,4,5-trisphosphate in the activation of protein kinase B

Author keywords

[No Author keywords available]

Indexed keywords

ADENOSINE TRIPHOSPHATE; CREATINE KINASE; CREATINE PHOSPHATE; MAGNESIUM CHLORIDE; ONCOPROTEIN; PHOSPHATIDYLINOSITOL; PHOSPHATIDYLINOSITOL 3 KINASE; PHOSPHATIDYLINOSITOL 3,4,5 TRISPHOSPHATE; PLECKSTRIN; PROTEIN KINASE B; THREONINE; UNCLASSIFIED DRUG;

EID: 0030799706     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5325.567     Document Type: Article
Times cited : (1080)

References (29)
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    • 2-terminal EEEEFMPME (Glu-Glu) tags (25) (E, Glu; F, Phe; M, Met; P, Pro).
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    • 3 and their isomers will be published separately.
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    • note
    • 32P]ATP was added to each assay. After affinity purification of the PKB, the beads were washed with TBS containing 1% NP-40 and 1 mM EDTA and then analyzed by SDS-polyacrylamide gel electrophoresis and autoradiography.
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    • note
    • There are at least two potential reasons why the two peaks of phosphorylation and activation reproducibly do not exactly coincide. The Glu-Glu-tagged PKB from Sf9 cells purifies as a heterogeneous mixture of phosphoproteins that is partially active. These species may exhibit differences in their ability to become phosphorylated versus their ability to become activated. An alternative possibility is that there is a second activity eluting earlier in the gradient that is required in addition to phosphorylation of PKB for activation, and it is when both this peak and the kinase peak overlap that the activation of PKB is seen. The two peak fractions were pooled for subsequent analysis of this activity.
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    • note
    • Radiolabeled peptides were sequenced in a Beckman 890C spinning cup sequenator in the presence of Polybrene and horse myoglobin. Each cycle fraction was blown dry in a chemical fume hood, redissolved in 6 ml of scintillation fluid and counted for 20 min, and corrected for background.
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    • note
    • This research is supported in part by the National Cancer Institute, NIH, Department of Health and Human Services under contract with ABL. P. Hawkins is a Biotechnology and Biological Sciences Research Council Research Fellow. We thank B. Burgering for the PSG5(HA)PKB, E. Porfiri for critical reading of the manuscript and helpful suggestions, D. Morrison for helpful suggestions, and Bayer Corporation for interactive support.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.