Sep 08 2005
Pál G, Fong SY, Kossiakoff AA, Sidhu SS
Protein Sci. 2005 Sep;14(9):2405-13
PMID: 16131663
Abstract
Combinatorial shotgun scanning mutagenesis was used to analyze two large, related protein binding sites to assess the specificity and importance of individual side chain contributions to binding affinity. The strategy allowed for cost-effective generation of a plethora of functional data. …
Jul 08 2005
Kouadio JL, Horn JR, Pal G, Kossiakoff AA
J. Biol. Chem. 2005 Jul;280(27):25524-32
PMID: 15857837
Abstract
The high affinity binding site (Site1) of the human growth hormone (hGH) binds to its cognate receptor (hGHR) via a concave surface patch containing about 35 residues. Using 167 sequences from a shotgun alanine scanning analysis of Site1, we …
Jun 08 2005
Bang D, Makhatadze GI, Tereshko V, Kossiakoff AA, Kent SB
Angew. Chem. Int. Ed. Engl. 2005 Jun;44(25):3852-6
PMID: 15834850
Abstract
Apr 08 2005
Pál G, Ultsch MH, Clark KP, Currell B, Kossiakoff AA, Sidhu SS
J. Mol. Biol. 2005 Apr;347(3):489-94
PMID: 15755445
Abstract
Combinatorial shotgun alanine-scanning was used to assess intramolecular cooperativity in the high affinity site (site 1) of human growth hormone (hGH) for binding to its receptor. A total of 19 side-chains were analyzed and statistically …
Jan 08 2005
Gopal B, Madan LL, Betz SF, Kossiakoff AA
Biochemistry 2005 Jan;44(1):193-201
PMID: 15628860
Abstract
Common structural motifs, such as the cupin domains, are found in enzymes performing different biochemical functions while retaining a similar active site configuration and structural scaffold. The soil bacterium Bacillus subtilis has 20 cupin genes (0.5% of the total genome) with …
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 …