Crystal structure of a bacterial albumin‐binding domain at 1.4 Å resolution

Cramer, Jacob Flyvholm; Nordberg, Peter; Hajdu, János; Lejon, Sara

doi:10.1016/j.febslet.2007.06.003

Cited by 14 publications

(13 citation statements)

References 16 publications

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“…GA (protein G-like albumin binding module) is found in a family of surface proteins of different bacterial species. It has been reported that the GA module binds close to a cleft at a site in domain IIA and domain IIIB of the albumin molecule 20,21 and involves a single site consisting of a segment spanning residues 330-548 ( Fig. 2D).…”

Section: Bacterial Protein and Albumin Associationmentioning

confidence: 95%

Ligand binding strategies of human serum albumin: How can the cargo be utilized?

et al. 2009

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Human serum albumin (HSA), being the most abundant carrier protein in blood and a modern day clinical tool for drug delivery, attracts high attention among biologists. Hence, its unfolding/refolding strategies and exogenous/endogenous ligand binding preference are of immense use in therapeutics and clinical biochemistry. Among its fellow proteins albumin is known to carry almost every small molecule. Thus, it is a potential contender for being a molecular cargo/or nanovehicle for clinical, biophysical and industrial purposes. Nonetheless, its structure and function are largely regulated by various chemical and physical factors to accommodate HSA to its functional purpose. This multifunctional protein also possesses enzymatic properties which may be used to convert prodrugs to active therapeutics. This review aims to highlight current overview on the binding strategies of protein to various ligands that may be expected to lead to significant clinical applications.

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Section: Bacterial Protein and Albumin Associationmentioning

confidence: 95%

Ligand binding strategies of human serum albumin: How can the cargo be utilized?

et al. 2009

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“…The ALB8 gene encoding the GA module was obtained as synthetic DNA in a carrier plasmid and was subcloned into a pET28 vector (Novagen) with an N-terminal histidine tag and thrombin protease cleavage site (HHHHSSGLVPRGSHM). The GA module was overexpressed and was then purified using an Ni 2+ -loaded chelating column and an imidazole gradient using standard procedures as described elsewhere (Cramer et al, 2007). Owing to the presence of heterogeneous cleavage products after thrombin digestion, the N-terminal tag was left intact for crystallization.…”

Section: Protein Purification Complex Formation and Crystallizationmentioning

confidence: 99%

Structural basis for the binding of naproxen to human serum albumin in the presence of fatty acids and the GA module

2008

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The previously determined crystal structure of the bacterial albumin‐binding GA module in complex with human serum albumin (HSA) suggested the possibility of utilizing the complex in the study of ligand binding to HSA. As a continuation of these studies, the crystal structure of the HSA–GA complex with the drug molecule naproxen and the fatty acid decanoate bound to HSA has been determined to a resolution of 2.5 Å. In terms of drug binding, the structure suggests that the binding of decanoate to the albumin molecule may play a role in making the haemin site in subdomain IB of the albumin molecule available for the binding of naproxen. In addition, structure comparisons with solved structures of HSA and of the HSA–GA complex show that the GA module is capable of binding to different conformations of HSA. The HSA–GA complex therefore emerges as a possible platform for the crystallographic study of specific HSA–drug interactions and of the influence exerted by the presence of fatty acids.

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“…This study demonstrated that the binding of G148-ABD to HSA can be abolished by only a few amino acid changes and the overall mapped binding region in G148-ABD is largely supported by NMR-perturbation studies performed on both the homologous ALB8-GA [26] and G148-ABD [4] and by the ALB8-GA:HSA structural complex [28]. However, the NMR-studies generally assign larger binding surfaces, which may in part be due to contacts between the albumin-binding domains, as indicated by the crystal structure of a dimer of ALB8-GA [27]. Neither NMR nor X-ray studies have specified the central importance of the second helix for binding as accurately as the mutational analysis of G148-ABD.…”

Section: Introductionmentioning

confidence: 99%

The Albumin-Binding Domain as a Scaffold for Protein Engineering

Nilvebrant

Hober

2013

Computational and Structural Biotechnology Journal

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The albumin-binding domain is a small, three-helical protein domain found in various surface proteins expressed by gram-positive bacteria. Albumin binding is important in bacterial pathogenesis and several homologous domains have been identified. Such albumin-binding regions have been used for protein purification or immobilization. Moreover, improvement of the pharmacokinetics, through the non-covalent association to albumin, by fusing such domains to therapeutic proteins has been shown to be successful. Domains derived from streptococcal protein G and protein PAB from Finegoldia magna, which share a common origin and therefore represent an interesting evolutionary system, have been thoroughly studied structurally and functionally. Their albumin-binding sites have been mapped and these domains form the basis for a wide range of protein engineering approaches. By substitution-mutagenesis they have been engineered to achieve a broader specificity, an increased stability or an improved binding affinity, respectively. Furthermore, novel binding sites have been incorporated either by replacing the original albumin-binding surface, or by complementing it with a novel interaction interface. Combinatorial protein libraries, where several residues have been randomized simultaneously, have generated a large number of new variants with desired binding characteristics. The albumin-binding domain has also been utilized to explore the relationship between three-dimensional structure and amino acid sequence. Proteins with latent structural information built into their sequence, where a single amino acid substitution shifts the equilibrium in favor of a different fold with a new function, have been designed. Altogether, these examples illustrate the versatility of the albumin-binding domain as a scaffold for protein engineering.

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Crystal structure of a bacterial albumin‐binding domain at 1.4 Å resolution

Cited by 14 publications

References 16 publications

Ligand binding strategies of human serum albumin: How can the cargo be utilized?

Ligand binding strategies of human serum albumin: How can the cargo be utilized?

Structural basis for the binding of naproxen to human serum albumin in the presence of fatty acids and the GA module

The Albumin-Binding Domain as a Scaffold for Protein Engineering

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