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G Dodson

Publications and source records attributed to G Dodson.

68 records · Page 4Linked to original sources
PubMed1987

An examination of the role of insulin dimerisation and negative cooperativity using the biological properties of the despentapeptide and deshexapeptide insulins.

The C-terminus of the insulin B chain is essential for dimerisation and expression of negative cooperativity. In order to evaluate the possible physiological role of these phenomena, we have studied the properties in vivo and in vitro of despentapeptide insulin (B26-30 deleted), derived from beef insulin, and deshexapeptide insulin (B25-30 deleted), derived from pork insulin. These materials do not dimerise and have 15% and 0% retention of negative cooperativity respective. Lipogenesis potencies in rat adipocytes were: despentapeptide insulin 19.9 +/- 0.3%; deshexapeptide insulin 19.9 +/- 1.5%. Binding potencies in adipocytes were: despentapeptide insulin 22.6 +/- 7.8%; deshexapeptide insulin 13.2 +/- 3.3%. Metabolic clearance rates were reduced compared to insulin (insulin = 19.1 +/- 0.9; despentapeptide insulin = 9.7 +/- 0.8; deshexapeptide insulin = 6.4 +/- 0.6 ml . min-1 . kg-1 at plasma concentration 0.5 nmol/l). Hypoglycaemic potencies were reduced for both analogues (40% and 30%) when calculated on the basis of plasma concentration although both analogues and insulin were equally effective at lowering plasma glucose concentration in equimolar doses. Plasma half-disappearance time was prolonged (despentapeptide insulin = 7.3 +/- 0.5; deshexapeptide insulin = 9.1 +/- 0.2 min). Both analogues were full agonists and conformed to the general relationship between in vitro and in vivo properties seen with a wide range of modified insulins. They resemble other analogues with modifications which reduce receptor affinity without impairing dimerisation or negative cooperativity. The results do not support a physiological role for dimerisation or negative cooperativity.

Adipose Tissue↗
PubMed1987

Resistance of the insulin crystal to lysosomal proteases: implications for pancreatic B-cell crinophagy.

Insulin is thought to be chemically stabilized within beta-granules in the crystal form. The other major products of the beta-granule, proinsulin and C-peptide, by contrast, are not thought able to crystallize. The physico-chemical properties of peptides in soluble or crystalline form are dramatically different. The ability of insulin to crystallize in the beta-granule might thus explain why this peptide, but not proinsulin/C-peptide, remains stable even after its introduction into lysosomes as occurs during granulolysis (crinophagy). We have now studied this by exposing proinsulin or insulin to lysosomal proteases in vitro. 125I-insulin in soluble form was found to be degraded at the same rate as 125I-proinsulin. Strikingly, however, when the labelled insulin was crystallized, its rate of degradation was decreased from 1.9 to 0.2 pmol/min. We take these data as confirmation that the insulin crystal is resistant to degradation, thereby possibly accounting for (a) the presence of insulin immunoreactivity within multigranular bodies, and (b) the unusually slow rate of degradation of insulin within B cells compared with that of other hormones in their cells of origin.

Animals↗
PubMed1986

Structure-function relationships of shortened [LeuB25]insulins, semisynthetic analogues of a mutant human insulin.

Replacement of B25-phenylalanine by leucine in the insulin sequence causes marked inactivation. The effect of this sequence variation was studied here in des-(B26-30)-insulin. [LeuB25]des-(B26-30)-insulin and its B25-amide were prepared by trypsin-mediated semisynthesis from N-terminally protected des-(B23-30)-insulin and synthetic tripeptides. The relative lipogenic potency in isolated rat adipocytes was 8.0% for the truncated analogue with a free B25-carboxyl function, and 18.1% for the amidated analogue. Binding to cultured human IM-9 lymphocytes was 4% and 9%, respectively. Thus, both shortened insulins are markedly more active than [LeuB25]insulin. The PheB25----LeuB25 substitution in both the shortened and the full sequence has a moderate effect on the CD spectrum, indicating that the gross main chain conformation is largely retained in both molecules. Independent of the substitution an absolute increase of the circular dichroism is observed upon amidation of the B25-carboxyl group.

Adipose Tissue↗
PubMed1983

Insulin.

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Amino Acid Sequence↗
PubMed1981

Evidence concerning insulin activity from the structure of a cross-linked derivative.

The important of crystallographic refinement for confident structural description, even at modest resolution, is demonstrated for N alpha A1,N epsilon B29-L,L-2,7-diaminosuberoyl (A2sb) insulin, a cross-linked insulin of low potency. The spatial arrangement of the cross-link itself can be described, and reliable estimates of the accuracy in atomic positions obtained. Comparison of invariant A2sb and native insulins shows a strong structural similarity, especially for the A chain surface residues and the dimer-forming residues of the B chain which have generally been strongly implicated in the receptor-binding region. Evidence from this analysis directs attention to the A chain, particularly the backbone, as being important in interactions with the membrane-bound receptor.

Animals↗
PubMedDate not supplied

Bonding of molecular oxygen to T state human haemoglobin.

Haemoglobin (Hb) is the tetrameric protein molecule that in vertebrate blood transports oxygen from the lungs to the tissues. This function depends on four subunits in the molecule binding cooperatively so that their affinity for oxygen increases as the level of oxygenation increases. X-ray analysis has shown that deoxyhaemoglobin, which has a low oxygen affinity, and oxyhaemoglobin, which has a high oxygen affinity, differ principally in their subunit or quaternary structures, referred to as the T and R states, respectively. As it switches from the T state to the R state during oxygenation, Hb increases its oxygen affinity. However, the structural pathway between deoxy- and oxy-haemoglobin is not known, principally because there has been no accurate structural knowledge of the intermediate states. We report here the crystal structure of T state human Hb in which the alpha chains are oxygenated and the beta subunits are oxygen-free. In this crystal the Hb appears to be in an intermediate state between the unliganded T state and the liganded R state. There is also evidence that the Hb molecule operates by loading and unloading the beta haems and thus the alpha-oxy, beta-deoxy Hb crystal may represent a physiologically important state.

Crystallography↗