Exploring and designing protein function with restricted diversity

Sidhu SS, Kossiakoff AA

Curr Opin Chem Biol 2007 Jun;11(3):347-54

PMID: 17500026


Combinatorial libraries with restricted diversity can be used to rapidly map binding energetics across protein interfaces. Shotgun scanning strategies have been used for alanine scanning and for alternative mutagenesis schemes that provide high-resolution functional views of binding interfaces. In addition, synthetic antibodies …

Continue reading

Time-controlled microfluidic seeding in nL-volume droplets to separate nucleation and growth stages of protein crystallization

Gerdts CJ, Tereshko V, Yadav MK, Dementieva I, Collart F, Joachimiak A, Stevens RC, Kuhn P, Kossiakoff A, Ismagilov RF

Angew. Chem. Int. Ed. Engl. 2006 Dec;45(48):8156-60

PMID: 17099920


Comprehensive and quantitative mapping of energy landscapes for protein-protein interactions by rapid combinatorial scanning

Pál G, Kouadio JL, Artis DR, Kossiakoff AA, Sidhu SS

J. Biol. Chem. 2006 Aug;281(31):22378-85

PMID: 16762925


A novel, quantitative saturation (QS) scanning strategy was developed to obtain a comprehensive data base of the structural and functional effects of all possible mutations across a large protein-protein interface. The QS scan approach was applied to …

Continue reading

The role of protein dynamics in increasing binding affinity for an engineered protein-protein interaction established by H/D exchange mass spectrometry

Horn JR, Kraybill B, Petro EJ, Coales SJ, Morrow JA, Hamuro Y, Kossiakoff AA

Biochemistry 2006 Jul;45(28):8488-98

PMID: 16834322


It is generally accepted that protein and solvation dynamics play fundamental roles in the mechanisms of protein-protein binding; however, assessing their contribution meaningfully has not been straightforward. Here, hydrogen/deuterium exchange mass spectrometry (H/D-Ex) was employed …

Continue reading

Crystal structure and site 1 binding energetics of human placental lactogen

Walsh ST, Kossiakoff AA

J. Mol. Biol. 2006 May;358(3):773-84

PMID: 16546209


In primates, placental lactogen (PL) is a pituitary hormone with fundamental roles during pregnancy involving fetal growth, metabolism, and stimulating lactation in the mother. Human placental lactogen (hPL) is highly conserved with human growth hormone (hGH) and both hormones bind to the hPRLR …

Continue reading

Dissecting the energetics of protein alpha-helix C-cap termination through chemical protein synthesis

Bang D, Gribenko AV, Tereshko V, Kossiakoff AA, Kent SB, Makhatadze GI

Nat. Chem. Biol. 2006 Mar;2(3):139-43

PMID: 16446709


The alpha-helix is a fundamental protein structural motif and is frequently terminated by a glycine residue. Explanations for the predominance of glycine at the C-cap terminal portions of alpha-helices have invoked uniquely favorable energetics of …

Continue reading

Structure of bistramide A-actin complex at a 1.35 angstroms resolution

Rizvi SA, Tereshko V, Kossiakoff AA, Kozmin SA

J. Am. Chem. Soc. 2006 Mar;128(12):3882-3

PMID: 16551075


Bistramide A is a highly potent antiproliferative marine natural product from Lissoclinum bistratum. We have previously established actin as the primary cellular receptor of bistramide A. We report herein the X-ray structure of bistramide A bound to monomeric …

Continue reading

The crystal structure of Aq_328 from the hyperthermophilic bacteria Aquifex aeolicus shows an ancestral histone fold

Qiu Y, Tereshko V, Kim Y, Zhang R, Collart F, Yousef M, Kossiakoff A, Joachimiak A

Proteins 2006 Jan;62(1):8-16

PMID: 16287087


The structure of Aq_328, an uncharacterized protein from hyperthermophilic bacteria Aquifex aeolicus, has been determined to 1.9 A by using multi-wavelength anomalous diffraction (MAD) phasing. Although the amino acid sequence analysis shows that …

Continue reading

Alternative views of functional protein binding epitopes obtained by combinatorial shotgun scanning mutagenesis

Pál G, Fong SY, Kossiakoff AA, Sidhu SS

Protein Sci. 2005 Sep;14(9):2405-13

PMID: 16131663


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. …

Continue reading

Shotgun alanine scanning shows that growth hormone can bind productively to its receptor through a drastically minimized interface

Kouadio JL, Horn JR, Pal G, Kossiakoff AA

J. Biol. Chem. 2005 Jul;280(27):25524-32

PMID: 15857837


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 …

Continue reading