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Structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling :The cases of calcineurin and protein kinase C iota.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling :
Reminder of title:	The cases of calcineurin and protein kinase C iota.
Author:	Roehrl, Michael Herbert Alexander.
Description:	282 p.
Notes:	Adviser: Gerhard Wagner.
Notes:	Source: Dissertation Abstracts International, Volume: 65-05, Section: B, page: 2404.
Contained By:	Dissertation Abstracts International65-05B.
Subject:	Chemistry, Biochemistry.
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3131970
ISBN:	0496792105

Structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling :The cases of calcineurin and protein kinase C iota.
Roehrl, Michael Herbert Alexander.

Structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling :The cases of calcineurin and protein kinase C iota.[electronic resource] - 282 p.

Adviser: Gerhard Wagner.

Thesis (Ph.D.)--Harvard University, 2004.

Section I focuses on the interaction between the phosphatase calcineurin and transcription factors of the NFAT family. We discovered novel small-molecule inhibitors of NFAT-calcineurin association (INCAs) that selectively disrupt this interaction. We introduce general methods for the rational development of competitive high-throughput screening assays for small-molecule inhibitors of protein-protein interactions by fluorescence polarization. INCAs do not compromise the enzymatic activity of calcineurin towards phosphorylated RII peptide, a substrate that does not require this NFAT docking site. In T cells, INCAs interfere selectively with the calcineurin-NFAT interaction. This substrate-selective enzyme inhibition represents a conceptual and practical advance over inhibition with drugs like cyclosporin A or FK506. Furthermore, we solved the 2.8-A crystal structure of the catalytic domain of human calcineurin A. Structural, mutational, and exhaustive computational INCA docking experiments lead us to a prediction of the VIVIT binding site near the catalytic domain's C-terminal beta sheet, but remote from its active site. We conclude with progress towards the complete backbone chemical shift assignment of the catalytic domain of calcineurin by multidimensional NMR spectroscopy.

ISBN: 0496792105Subjects--Topical Terms:

226900
Chemistry, Biochemistry.

Structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling :The cases of calcineurin and protein kinase C iota.
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245 1 0 $a Structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling : $b The cases of calcineurin and protein kinase C iota. $h [electronic resource]
300 $a 282 p.
500 $a Adviser: Gerhard Wagner.
500 $a Source: Dissertation Abstracts International, Volume: 65-05, Section: B, page: 2404.
502 $a Thesis (Ph.D.)--Harvard University, 2004.
520 # $a Section I focuses on the interaction between the phosphatase calcineurin and transcription factors of the NFAT family. We discovered novel small-molecule inhibitors of NFAT-calcineurin association (INCAs) that selectively disrupt this interaction. We introduce general methods for the rational development of competitive high-throughput screening assays for small-molecule inhibitors of protein-protein interactions by fluorescence polarization. INCAs do not compromise the enzymatic activity of calcineurin towards phosphorylated RII peptide, a substrate that does not require this NFAT docking site. In T cells, INCAs interfere selectively with the calcineurin-NFAT interaction. This substrate-selective enzyme inhibition represents a conceptual and practical advance over inhibition with drugs like cyclosporin A or FK506. Furthermore, we solved the 2.8-A crystal structure of the catalytic domain of human calcineurin A. Structural, mutational, and exhaustive computational INCA docking experiments lead us to a prediction of the VIVIT binding site near the catalytic domain's C-terminal beta sheet, but remote from its active site. We conclude with progress towards the complete backbone chemical shift assignment of the catalytic domain of calcineurin by multidimensional NMR spectroscopy.
520 # $a Section II describes the three-dimensional structure of the V1 domain of human protein kinase C iota (PKC &igr;) by NMR spectroscopy. Dysregulation of PKC &igr; makes leukemia and breast cancer cells highly resistant to chemotherapeutic agents. The N-terminal V1 domain of PKC &igr; participates in cellular targeting and regulation, thus rendering it an attractive object for pharmacologic intervention. We describe the biosynthetic production and biochemical purification of recombinant PKC &igr; V1. Subsequently, the chemical shift assignment and the solution structure of the V1 domain were obtained using NMR spectroscopy. The protein assumes an ubiquitin-like beta grasp fold. We close with an outlook on work that investigates the interaction of PKC &igr; V1 with Par-6.
520 # $a The Appendix addresses an interesting, but often overlooked, issue concerning the correspondence between spin-dynamic and pulse program phase behavior of NMR spectrometers. The results are important for spin state selective experiments, such as TROSY-type experiments, which I have implemented and relied on for the NMR spectroscopy of calcineurin.
520 # $a This thesis describes structural and chemical genetic insights into protein-protein interactions of phosphate-dependent cell signaling.
590 $a School code: 0084.
650 # 0 $a Chemistry, Biochemistry. $3 226900
650 # 0 $a Biophysics, General. $3 226901
650 # 0 $a Health Sciences, Immunology. $3 226966
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690 $a 0786
690 $a 0982
710 0 # $a Harvard University. $3 212445
773 0 # $g 65-05B. $t Dissertation Abstracts International
790 $a 0084
790 1 0 $a Wagner, Gerhard, $e advisor
791 $a Ph.D.
792 $a 2004
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