Racemic crystallography of synthetic protein enantiomers used to determine the X-ray structure of plectasin by direct methods

Mandal K, Pentelute BL, Tereshko V, Thammavongsa V, Schneewind O, Kossiakoff AA, Kent SB

Protein Sci. 2009 Jun;18(6):1146-54

PMID: 19472324


We describe the use of racemic crystallography to determine the X-ray structure of the natural product plectasin, a potent antimicrobial protein recently isolated from fungus. The protein enantiomers L-plectasin and D-plectasin were prepared by …

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Crystal structure of full-length KcsA in its closed conformation

Uysal S, Vásquez V, Tereshko V, Esaki K, Fellouse FA, Sidhu SS, Koide S, Perozo E, Kossiakoff A

Proc. Natl. Acad. Sci. U.S.A. 2009 Apr;106(16):6644-9

PMID: 19346472


KcsA is a proton-activated, voltage-modulated K(+) channel that has served as the archetype pore domain in the Kv channel superfamily. Here, we have used synthetic antigen-binding fragments …

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X-ray structure of native scorpion toxin BmBKTx1 by racemic protein crystallography using direct methods

Mandal K, Pentelute BL, Tereshko V, Kossiakoff AA, Kent SB

J. Am. Chem. Soc. 2009 Feb;131(4):1362-3

PMID: 19133782


Racemic protein crystallography, enabled by total chemical synthesis, has allowed us to determine the X-ray structure of native scorpion toxin BmBKTx1; direct methods were used for phase determination. This is the first example of a protein …

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Principal determinants leading to transition state formation of a protein-protein complex, orientation trumps side-chain interactions

Horn JR, Sosnick TR, Kossiakoff AA

Proc. Natl. Acad. Sci. U.S.A. 2009 Feb;106(8):2559-64

PMID: 19196954


The binding transition state (TS) is the rate-limiting step for transient molecular interactions. This important step in the molecular recognition process, however, is largely understood only at a qualitative level. To establish a more quantitative picture of the TS …

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Understanding mechanisms governing protein-protein interactions from synthetic binding interfaces

Kossiakoff AA, Koide S

Curr. Opin. Struct. Biol. 2008 Aug;18(4):499-506

PMID: 18638552


Recent advances in methodologies and design of combinatorial library selection have enabled comprehensive characterization of sequence space for protein-protein interaction interfaces and generation of fully synthetic binding interfaces. By exhaustively introducing and quantitatively analyzing mutations in natural interfaces, new insights into their …

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Toward chaperone-assisted crystallography: protein engineering enhancement of crystal packing and X-ray phasing capabilities of a camelid single-domain antibody (VHH) scaffold

Tereshko V, Uysal S, Koide A, Margalef K, Koide S, Kossiakoff AA

Protein Sci. 2008 Jul;17(7):1175-87

PMID: 18445622


A crystallization chaperone is an auxiliary protein that binds to a target of interest, enhances and modulates crystal packing, and provides high-quality phasing information. We critically evaluated the effectiveness of a camelid single-domain antibody (V(H)H) as …

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X-ray structure of snow flea antifreeze protein determined by racemic crystallization of synthetic protein enantiomers

Pentelute BL, Gates ZP, Tereshko V, Dashnau JL, Vanderkooi JM, Kossiakoff AA, Kent SB

J. Am. Chem. Soc. 2008 Jul;130(30):9695-701

PMID: 18598029


Chemical protein synthesis and racemic protein crystallization were used to determine the X-ray structure of the snow flea antifreeze protein (sfAFP). Crystal formation from a racemic solution containing equal amounts of the …

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The 1.38 A crystal structure of DmsD protein from Salmonella typhimurium, a proofreading chaperone on the Tat pathway

Qiu Y, Zhang R, Binkowski TA, Tereshko V, Joachimiak A, Kossiakoff A

Proteins 2008 May;71(2):525-33

PMID: 18175314


The DmsD protein is necessary for the biogenesis of dimethyl sulphoxide (DMSO) reductase in many prokaryotes. It performs a critical chaperone function initiated through its binding to the twin-arginine signal peptide of DmsA, the catalytic subunit of …

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Synthetic antibodies for specific recognition and crystallization of structured RNA

Ye JD, Tereshko V, Frederiksen JK, Koide A, Fellouse FA, Sidhu SS, Koide S, Kossiakoff AA, Piccirilli JA

Proc. Natl. Acad. Sci. U.S.A. 2008 Jan;105(1):82-7

PMID: 18162543


Antibodies that bind protein antigens are indispensable in biochemical research and modern medicine. However, knowledge of RNA-binding antibodies and their application in the ever-growing RNA field is …

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High-throughput generation of synthetic antibodies from highly functional minimalist phage-displayed libraries

Fellouse FA, Esaki K, Birtalan S, Raptis D, Cancasci VJ, Koide A, Jhurani P, Vasser M, Wiesmann C, Kossiakoff AA, Koide S, Sidhu SS

J. Mol. Biol. 2007 Nov;373(4):924-40

PMID: 17825836


We have previously established a minimalist approach to antibody engineering by using a phage-displayed framework to support complementarity determining region (CDR) diversity restricted …

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