Dec 08 2004
Walsh ST, Sylvester JE, Kossiakoff AA
Proc. Natl. Acad. Sci. U.S.A. 2004 Dec;101(49):17078-83
PMID: 15563602
Abstract
Growth hormone regulates its biological properties via a sequential hormone-induced receptor homodimerization mechanism. Using a mutagenesis-scanning analysis of 81 single and 32 pairwise double mutations, we show that the hormone’s two spatially distal receptor binding sites (Site1 and Site2) …
Jun 08 2004
Kossiakoff AA
Adv. Protein Chem. 2004;68:147-69
PMID: 15500861
Abstract
The pituitary hormones growth hormone (GH), prolactin (PRL) and placental lactogen (PL), are members of an extensive cytokine superfamily of hormones and receptors that share many of the same general structure-function relationships in expressing their biological activities. The biology of the pituitary hormones involves a very …
May 08 2004
Bernat B, Sun M, Dwyer M, Feldkamp M, Kossiakoff AA
Biochemistry 2004 May;43(20):6076-84
PMID: 15147191
Abstract
Phage display mutagenesis is a widely used approach to engineering novel protein properties and is especially powerful in probing structure-function relationships in molecular recognition processes. The relative contributions of additive and cooperative binding forces and the influence of conformational …
Sep 08 2003
Walsh ST, Jevitts LM, Sylvester JE, Kossiakoff AA
Protein Sci. 2003 Sep;12(9):1960-70
PMID: 12930995
Abstract
Receptor signaling in the growth hormone (GH)-growth hormone receptor (GHR) system is controlled through a sequential two-step hormone-induced dimerization of two copies of the extracellular domain (ECD) of the receptor. The regulatory step of this process is the binding of …
Sep 08 2003
Pal G, Kossiakoff AA, Sidhu SS
J. Mol. Biol. 2003 Sep;332(1):195-204
PMID: 12946357
Abstract
A high-affinity variant of human growth hormone (hGH(v)) contains 15 mutations within site 1 and binds to the hGH receptor (hGHR) approximately 400-fold tighter than does wild-type (wt) hGH (hGH(wt)). We used shotgun scanning combinatorial mutagenesis to dissect the energetic contributions …
Jun 08 2003
Pál G, Santamaria F, Kossiakoff AA, Lu W
Protein Expr. Purif. 2003 Jun;29(2):185-92
PMID: 12767808
Abstract
Selective incorporation of non-natural amino acid residues into proteins is a powerful approach to delineate structure-function relationships. Although many methodologies are available for chemistry-based protein engineering, more facile methods are needed to make this approach suitable for routine laboratory …
Feb 08 2003
Bernat B, Pal G, Sun M, Kossiakoff AA
Proc. Natl. Acad. Sci. U.S.A. 2003 Feb;100(3):952-7
PMID: 12552121
Abstract
Signaling in the human growth hormone (hGH)-human GH receptor system is initiated by a controlled sequential two-step hormone-induced dimerization of two hGH receptors via their extracellular domains (ECDs). Little is currently known about the energetics governing the …
Feb 08 2002
Schiffer C, Ultsch M, Walsh S, Somers W, de Vos AM, Kossiakoff A
J. Mol. Biol. 2002 Feb;316(2):277-89
PMID: 11851338
Abstract
The structure of the ternary complex between the phage display- optimized, high-affinity Site 1 variant of human growth hormone (hGH) and two copies of the extracellular domain (ECD) of the hGH receptor (hGHR) has …
Nov 08 2001
Dwyer JJ, Dwyer MA, Kossiakoff AA
Biochemistry 2001 Nov;40(45):13491-500
PMID: 11695896
Abstract
Using phage display mutagenesis, high affinity variants of RNase S-peptide were produced that bind to RNase S-protein over 100-fold more tightly than the wild type S-peptide. The S-peptide: S-protein interface was further characterized using “biased” phage display libraries, where each targeted residue was …
Apr 08 2000
Dwyer MA, Lu W, Dwyer JJ, Kossiakoff AA
Chem. Biol. 2000 Apr;7(4):263-74
PMID: 10780926
Abstract
BACKGROUND: Molecular diversity in nature is developed through a combination of genetic and chemical elements. We have developed a method that permits selective manipulation of both these elements in one protein engineering tool. It combines the ability to introduce non-natural …