Keyhole Limpet Hemocyanin: 9-Å CryoEM Structure and Molecular Model of the KLH1 Didecamer Reveal the Interfaces and Intricate Topology of the 160 Functional Units

Gatsogiannis, Christos; Markl, Jürgen

doi:10.1016/j.jmb.2008.10.080

Cited by 84 publications

(128 citation statements)

References 62 publications

Supporting

Mentioning

118

Contrasting

Order By: Relevance

“…This was, especially, straightforward, because the morphological unit of FU-h in the crystal is a dimer, which, according to the docking results, exactly corresponds to the FU-h dimer naturally occurring in the KLH1 molecule ( Fig. 2) (20).…”

Section: Introductionmentioning

confidence: 68%

“…The resulting structure could be stably docked, by automated rigid-body fitting, to a 9 Å cryo-electron microscopical structure of the KLH1 decamer, which revealed the architecture of the outer collar (20). This was, especially, straightforward, because the morphological unit of FU-h in the crystal is a dimer, which, according to the docking results, exactly corresponds to the FU-h dimer naturally occurring in the KLH1 molecule ( Fig.…”

Section: Introductionmentioning

confidence: 83%

“…Briefly, KLH1-h was prepared from purified KLH1 by limited proteolysis with V8 protease and subsequently purified by ion exchange chromatography (20,26). KLH1-h was then crystallized at pH 4.0 in sodium citrate buffer in the presence of 9.0% PEG 1000 and 50-mM K 2 HPO 4 .…”

Section: Previous Structure Determinationmentioning

confidence: 99%

“…The only exception to the standard architecture of mollusc FUs is FU-h, which forms the outer collar of the decamer (20). Unlike all other FUs, FU-h has an additional C-terminal extension of approximately 100 amino acids, which marks the very end of the molluscan hemocyanin subunit polypeptide (21).…”

Section: Introductionmentioning

confidence: 99%

“…An offset of one or two amino acids could not be excluded, notably in the cupredoxin-like domain in which the sequence similarity with the template (cucumber basic protein) was very low (6). Consequently, in the description of the putative inter-FU contacts within the KLH1 molecule, the amino acid side chains of the cupredoxin-like domain were not considered (20).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

The refined structure of functional unit h of keyhole limpet hemocyanin (KLH1‐h) reveals disulfide bridges

et al. 2011

Self Cite

View full text Add to dashboard Cite

Hemocyanins are multimeric oxygen‐transport proteins in the hemolymph of many arthropods and mollusks. The overall molecular architecture of arthropod and molluscan hemocyanin is very different, although they possess a similar binuclear type 3 copper center to bind oxygen in a side‐on conformation. Gastropod hemocyanin is a 35 nm cylindrical didecamer (2 × 10‐mer) based on a 400 kDa subunit. The latter is subdivided into eight paralogous “functional units” (FU‐a to FU‐h), each with an active site. FU‐a to FU‐f contribute to the cylinder wall, whereas FU‐g and FU‐h form the internal collar complex. Atomic structures of FU‐e and FU‐g, and a 9 Å cryoEM structure of the 8 MDa didecamer are available. Recently, the structure of keyhole limpet hemocyanin FU‐h (KLH1‐h) was presented as a Cα‐trace at 4 Å resolution. Unlike the other seven FU types, FU‐h contains an additional C‐terminal domain with a cupredoxin‐like fold. Because of the resolution limit of 4 Å, in some loops, the course of the protein backbone could not be established with high certainty yet. Here, we present a refined atomic structure of FU‐h (KLH1‐h) obtained from low‐resolution refinement, which unambiguously establishes the course of the polypeptide backbone and reveals the disulfide bridges as well as the orientation of bulky amino acids. © 2011 IUBMB IUBMB Life, 63(3): 183–187, 2011

show abstract

Section: Introductionmentioning

confidence: 68%

Section: Introductionmentioning

confidence: 83%

Section: Previous Structure Determinationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The refined structure of functional unit h of keyhole limpet hemocyanin (KLH1‐h) reveals disulfide bridges

et al. 2011

Self Cite

View full text Add to dashboard Cite

show abstract

Structures, Functions, and Assembly of Tyrosinases and Hemocyanins

Fujieda

Itoh

2017

Encyclopedia of Inorganic and Bioinorganic Chemistry

View full text Add to dashboard Cite

Metal‐assembling processes in the metalloproteins and metalloenzymes have attracted much recent attention due to their strong relevance not only to enzymology but also to medical science. Concerning the coupled dinuclear copper proteins, so‐called type‐3 copper proteins, a growing amount of information has been accumulated during the past decades. In this chapter, we introduce how copper ions are sorted and transferred to the type‐3 copper proteins in prokaryotic as well as eukaryotic cells. Furthermore, the detailed molecular mechanisms of copper incorporation processes in the type‐3 copper protein active sites will be depicted especially in the bacterial and fungal tyrosinases.

show abstract

Cellular and Biomolecular Recognition: Synthetic and non‐Biological Molecules. Edited by Raz Jelinek.

Thomas¹

2010

ChemBioChem

View full text Add to dashboard Cite

review the use of immobilized ligands for proteomic applications (affinity chromatography, chemical microarrays), including an excellent compilation of case studies in which the successful identification of the protein target of a bioactive small molecule illuminated the underlying biology. This chapter also provides numerous examples for reactive proteomics. A more in-depth coverage of this blossoming field, sometimes also called activity-based protein profiling, would have been well within the scope of this book.Four more specialized chapters highlight selected applications of small molecule-protein interactions, including an excellent review on fluorescent probes for the in vivo detection of metabolites. Chapter 8 covers the medicinal chemistry of nuclear hormone receptor ligands and their successful translation into the clinic. Chapter 9 emphasizes the impact of small-molecule inhibitors on the understanding of cell motility. Chapter 10 outlines the metabolic engineering of glycoproteins and their subsequent modification with small-molecule probes.The individual chapters are generally well structured and well supported by references for further reading. The quality of the graphics is adequate, with additional colour figures where necessary. Since most chapters present more advanced techniques and/or specialized applications in chemical biology, a sound background in organic chemistry and biochemistry is absolutely required. Given the sheer breadth and diversity of topics that have developed at the interface between chemistry and biology in the past decade, Protein Targeting with Small Molecules does not claim to cover all aspects of chemical biology.Rather, it provides a concise and focused overview centred on how to address small molecule-protein interactions.As such, the book is well suited for advanced graduate students already dedicated to the field or for researchers from related disciplines who want to expand their background at the interface between chemistry and biology. It will be especially valuable to scientists with an interest in translational research and in the prospects of chemical biology towards drug discovery.

show abstract

Keyhole Limpet Hemocyanin: 9-Å CryoEM Structure and Molecular Model of the KLH1 Didecamer Reveal the Interfaces and Intricate Topology of the 160 Functional Units

Cited by 84 publications

References 62 publications

The refined structure of functional unit h of keyhole limpet hemocyanin (KLH1‐h) reveals disulfide bridges

The refined structure of functional unit h of keyhole limpet hemocyanin (KLH1‐h) reveals disulfide bridges

Structures, Functions, and Assembly of Tyrosinases and Hemocyanins

Cellular and Biomolecular Recognition: Synthetic and non‐Biological Molecules. Edited by Raz Jelinek.

Contact Info

Product

Resources

About