Insulin: The Structure in the Crystal and its Reflection in Chemistry and Biology by

Blundell, Tom L.; Dodson, Guy; Hodgkin, Dorothy Crowfoot; Mercola, Dan

doi:10.1016/s0065-3233(08)60143-6

Cited by 832 publications

(612 citation statements)

References 200 publications

Supporting

Mentioning

574

Contrasting

Unclassified

Order By: Relevance

“…With the advent of recombinant DNA technology, single amino acid substitutions could be targeted to regions involved in hexamer formation that also lie on the periphery of the insulin receptor (IR) binding regions [10]. The histidine residue at the B10 position was known to be involved in self-association through the coordination of zinc ions by the imidazole side chains [13]. One of the analogues described by Brange and colleagues [10] replaced this histidine residue with the negatively charged aspartic acid, creating the insulin analogue known as insulin X10.…”

Section: The Development Of Insulin X10mentioning

confidence: 99%

Insulin X10 revisited: a super-mitogenic insulin analogue

et al. 2011

View full text Add to dashboard Cite

The molecular safety of insulin analogues has received a great deal of attention over the last year. In particular, attention has been directed to the mitogenic properties of insulin analogues as compared with human insulin. Understanding the mechanisms implicated in mediating mitogenic effects of insulin is therefore of particular interest. In this review we detail the story of the rapid-acting insulin analogue known as X10, which was the first insulin analogue in clinical development, but ended up being discontinued at an early clinical development stage following findings of mammary tumours in female Sprague-Dawley rats. The molecular characteristics of insulin X10, along with its interaction at both the IGF-1 receptor and the insulin receptor, have provided us with important insights into mechanisms implicated in metabolic and mitogenic signalling of insulin analogues.

show abstract

Section: The Development Of Insulin X10mentioning

confidence: 99%

Insulin X10 revisited: a super-mitogenic insulin analogue

et al. 2011

View full text Add to dashboard Cite

show abstract

“…5). The slow reaction with the second interchain bridge may perhaps be attributed to its being more highly buried and in a more hydrophobic environment than either of the other bonds (Blundell et al, 1972). In another context an attempt was made to use TCEP for removing a t-butylthio protecting group from synthetic peptides.…”

Section: Selectivity and Kinetics Of Reductionmentioning

confidence: 99%

Disulfide structures of highly bridged peptides: A new strategy for analysis

1993

View full text Add to dashboard Cite

A new approach is described for analyzing disulfide linkage patterns in peptides containing tightly clustered cystines. Such peptides are very difficult to analyze with traditional strategies, which require that the peptide Chain be split between close or adjacent Cys residues. The water-soluble tris-(2-~arboxyethyl)-phosphine (TCEP) reduced disulfides at pH 3, and partially reduced peptides were purified by high performance liquid chromatography with minimal thiol-disulfide exchange. Alkylation of free thiols, followed by sequencer analysis, provided explicit assignment of disulfides that had been reduced. Thiol-disulfide exchange occurred during alkylation of some peptides, but correct deductions were still possible. Alkylation competed best with exchange when peptide solution was added with rapid mixing to 2.2 M iodoacetamide. Variants were developed in which up to three alkylating agents were used to label different pairs of thiols, allowing a full assignment in one sequencer analysis. Model peptides used included insulin (three bridges, intra-and interchain disulfides; -Cys.Cys-pair), endothelin and apamin (two disulfides; -Cys.x.Cys-pair), conotoxin GI and isomers (two disulfides; -Cys.Cys-pair), and bacterial enterotoxin (three bridges within 13 residues; two -Cys.Cys-pairs). With insulin, all intermediates in the reduction pathway were identified; with conotoxin GI, analysis was carried out successfully for all three disulfide isomers. In addition to these known structures, the method has been applied successfully to the analysis of several previously unsolved structures of similar complexity. Rates of reduction of disulfide bonds varied widely, but most peptides did not show a strongly preferred route for reduction.

show abstract

“…For human insulin, the presence of zinc promotes association to the hexameric state in what is referred to as the Testate conformation, where two zinc ions are bound by the HisB" residues of each subunit. An octahedral coordination geometry is completed by three water molecules (Blundell et al, 1972;Baker et al, 1988). The addition of phenolic ligands induces a conformational transition by binding to specific sites on the insulin hexamer, resulting in the N-terminal eight amino acids of the B-chain converting from an extended conformation to an a-helix.…”

Section: Stability and Conformution Of Ligand-bound Lyspro Hexamermentioning

confidence: 99%

“…The hexamer is mainly stabilized through zinc coordination, but additional polar and nonpolar residues are buried between the dimers as a result of hexamer assembly (Baker et al, 1988). The predominant nonpolar dimer contacts involve the C-terminal end of the B-chain, with B23-B26 and B28 being the most significant (Blundell et al, 1972;Baker et al, 1988). Association of the dimer is secured by the small antiparallel P-sheet of hydrogen bonds involving residues B24 and B26 (Baker et al, 1988).…”

mentioning

confidence: 99%

Physicochemical basis for the rapid time‐action of Lys^B28Pro^B29‐insulin: Dissociation of a protein‐ligand complex

et al. 1996

View full text Add to dashboard Cite

The rate-limiting step for the absorption of insulin solutions after subcutaneous injection is considered to be the dissociation of self-associated hexamers to monomers. To accelerate this absorption process, insulin analogues have been designed that possess full biological activity and yet have greatly diminished tendencies to self-associate. Sedimentation velocity and static light scattering results show that the presence of zinc and phenolic ligands (m-cresol and/or phenol) cause one such insulin analogue, L y~~~~P r o~~~-h u m a n insulin (LysPro), to associate into a hexameric complex. Most importantly, this ligand-bound hexamer retains its rapid-acting pharmacokinetics and pharmacodynamics. The dissociation of the stabilized hexameric analogue has been studied in vitro using static light scattering as well as in vivo using a female pig pharmacodynamic model. Retention of rapid time-action is hypothesized to be due to altered subunit packing within the hexamer. Evidence for modified monomer-monomer interactions has been observed in the X-ray crystal structure of a zinc LysPro hexamer (Ciszak E et al., 1995, Structure 3:615-622). The solution state behavior of LysPro, reported here, has been interpreted with respect to the crystal structure results. In addition, the phenolic ligand binding differences between LysPro and insulin have been compared using isothermal titrating calorimetry and visible absorption spectroscopy of cobalt-containing hexamers. These studies establish that rapid-acting insulin analogues of this type can be stabilized in solution via the formation of hexamer complexes with altered dissociation properties.

show abstract

Insulin: The Structure in the Crystal and its Reflection in Chemistry and Biology by

Cited by 832 publications

References 200 publications

Insulin X10 revisited: a super-mitogenic insulin analogue

Insulin X10 revisited: a super-mitogenic insulin analogue

Disulfide structures of highly bridged peptides: A new strategy for analysis

Physicochemical basis for the rapid time‐action of Lys^B28Pro^B29‐insulin: Dissociation of a protein‐ligand complex

Contact Info

Product

Resources

About

Insulin: The Structure in the Crystal and its Reflection in Chemistry and Biology by

Cited by 832 publications

References 200 publications

Insulin X10 revisited: a super-mitogenic insulin analogue

Insulin X10 revisited: a super-mitogenic insulin analogue

Disulfide structures of highly bridged peptides: A new strategy for analysis

Physicochemical basis for the rapid time‐action of LysB28ProB29‐insulin: Dissociation of a protein‐ligand complex

Contact Info

Product

Resources

About

Physicochemical basis for the rapid time‐action of Lys^B28Pro^B29‐insulin: Dissociation of a protein‐ligand complex