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Biomedical subjects

G E Chapman

Publications and source records attributed to G E Chapman.

At least 37 records · Page 2Linked to original sources
PubMed1987

Methionine oxidation in human growth hormone and human chorionic somatomammotropin. Effects on receptor binding and biological activities.

The oxidation of the methionine residues of human growth hormone (hGH) and human chorionic somatomammotropin (hCS) to methionine sulfoxide by hydrogen peroxide has been studied. The kinetics of oxidation of individual methionine residues has been measured by reverse-phase high pressure liquid chromatography tryptic peptide mapping. Met-170 is completely resistant to oxidation in both hormones. The other 3 methionine residues in hCS (Met-64, Met-96, and Met-179) have markedly different reaction rates. Oxidation of the methionine residues does not appear to cause gross conformational changes in either hGH or hCS, as judged by CD and 1H NMR spectroscopy. Oxidation of Met-14 and Met-125 in hGH has little effect on affinity of the hormone for lactogenic receptors or on its potency in the Nb2 rat lymphoma in vitro bioassay for lactogenic hormones. The oxidation of Met-64 and/or Met-179 in hCS reduces profoundly both its affinity for lactogenic receptors and its in vitro biological potency. It is inferred by induction that residues 64 and/or 179 are critical for the binding of both hGH and hCS to lactogenic receptors and the expression of lactogenic biological activity.

Animals↗
PubMed1987

The amino acid sequence of Emu osteocalcin: gas phase sequencing of Gla-containing proteins.

Osteocalcin (OC), the major gamma carboxyglutamic acid (Gla)-containing protein of vertebrate bone, has been isolated from bones of the emu (Dromaius novaehollandae) and the primary structure determined by a combination of gas phase N-terminal sequencing of the intact molecule and a proteolytic fragment, and carboxypeptidase Y C-terminal sequencing. Gla residues were located by counting tritium radioactivity in fractions from the N-terminal sequencing of the tritiated/thermally decarboxylated molecule. Emu OC consists of 48 amino acid residues containing 3 Gla residues, and a single disulphide bond. The C-terminal 29 residues are identical to those of the human and sheep OC sequences. Alignment of the N-terminal sequence against those of other OCs reveals greater sequence homology with chicken OC than with mammalian OCs.

1-Carboxyglutamic Acid↗
PubMed1986

Low serum osteocalcin levels in glucocorticoid-treated asthmatics.

Serum osteocalcin (OC) levels were measured in 19 asthmatic patients receiving long term glucocorticoid therapy and in age- and sex-matched asthmatic patients not receiving this treatment. In the glucocorticoid-treated patients, the mean OC level was approximately 50% less than that in the control group (P less than 0.001), and there was a direct correlation between serum OC and 1,25-dihydroxyvitamin D [1,25-(OH)2D; r = 0.71; P less than 0.001]. Multiple regression analysis in a total of 39 glucocorticoid-treated patients indicated that OC correlated directly to 1,25-(OH)2D and inversely to glucocorticoid dose. There was no correlation between OC and 1,25-(OH)2D in the control group and no significant difference in mean serum 1,25-(OH)2D between the steroid-treated asthmatic patients and the asthmatic control patients. The effect of a 4-day course of oral 1,25-(OH)2D on serum OC was studied in six patients with glucocorticoid excess and six normal subjects. There was a similar percent increase in OC levels in both groups, though the basal concentrations and absolute increases were substantially less in the steroid-treated group. It is likely that the depression of serum OC in glucocorticoid-treated patients results from the reduction in the rate of bone formation induced by these hormones.

Administration, Oral↗
PubMed1985

Immunochemical detection and characterisation of osteocalcin from moa bone.

Osteocalcin (the 6,000 dalton Mr gamma-carboxyglutamate-containing protein of bone) has been detected in acid extracts of bones of the extinct class of New Zealand ratite birds, the moas, using a radioimmunoassay for sheep osteocalcin. The immunoreactive osteocalcin of the extracts of two of these bones (the fibulae from two specimens of Pachyornis elephantopus found in South Island swamps) has been fractionated by gel filtration chromatography and reversed-phase high performance liquid chromatography, and behaves in a manner characteristic of osteocalcin from modern bones. Carbon-14 dating of bones and gizzard contents found in association with these specimens indicates approximate ages of 3,600 and 7,400 years respectively.

Animals↗
PubMed1985

Population studies: an integrated course in epidemiology and sociology for medical students.

At the Royal Free Hospital School of Medicine, London, an integrated course in epidemiology and sociology for preclinical students was introduced in 1979. The course--Population Studies--is taken by the 100 second-year medical students in the summer term before they enter their clinical years. It occupies one full day and one half day each week for 8 weeks--approximately 80 hours of tuition. Population Studies is unusual in two respects. Firstly, it introduces a substantial amount of epidemiology into the preclinical curriculum. And, secondly, this is the only London medical school to integrate the teaching of sociology and epidemiology into the one course.

Education, Medical, Undergraduate↗
PubMed1984

The amino acid sequences of goat, pig and wallaby osteocalcins.

The amino acid sequences of osteocalcins from cortical long bones of goat, pig and wallaby have been determined by manual microsequencing methods. The location of the gamma-carboxyglutamic acid residues were identified by N-terminal degradation of peptides in which these residues had been tritium-labelled in the gamma-H position and subsequently thermally decarboxylated to glutamic acid residues. Glutamic-17 is not gamma-carboxylated in wallaby osteocalcin, unlike other osteocalcins so far sequenced (except human). This may be related to sequence changes in the flanking residues. Proline-9 is fully hydroxylated in wallaby osteocalcin, despite the sequence substitution Ala-8----Phe-8.

Amino Acid Sequence↗
PubMed1983

A proton-nuclear-magnetic-resonance study of human somatotropin (growth hormone). Assignment and properties of the histidine residues.

The 1H-n.m.r. spectra of human somatotropin (growth hormone) show perturbed peaks from individual aromatic and aliphatic apolar residues, characteristic of a specifically folded globular structure. The imidazole C-2-H resonances of the histidine residues (at positions 18, 21 and 151 in the somatotropin sequence) were individually resolved, and their titration behaviour in the pH range 1.2-11.5 was investigated. The imidazole C-2-H resonance of histidine-151 is assigned, by comparison of its titration behaviour in human somatotropin and desamido-somatotropin (Asn-152 leads to Asp-152). The C-2-H resonances of all three histidine residues are assigned, by comparison of their relative deuterium-exchange rates (determined by n.m.r.) and the relative tritium-exchange rates of the histidine residues (determined by tryptic digestion of tritiated human somatotropin and reversed-phase high-pressure liquid-chromatographic separation of the histidine-containing tryptic peptides). There is evidence that histidine-18 forms an ion-pair bond with a glutamic acid or aspartic acid residue. The globular structure does not appear to change from pH3 to 11.5, though there is evidence for an unfolding of a region of the structure (involving histidine-21 and a tyrosine residue) below pH3.

Chemical Phenomena↗
PubMed1983

Ritual and rational action in hospitals.

Menzies argues that nursing hierarchies and ritual practices protect nurses from the anxieties provoked by encountering human suffering. This proposition is examined with particular reference to ritual practices in nursing. It is argued that Menzies studied nurses in isolation from the societal and subcultural norms and values which direct hospital activity. Her psychodynamic model is contrasted with a sociological model of human conduct and action. The characteristics of ritual and rational action, and the difference between non-rational and irrational rituals, is explored. The findings of three 5-month periods of participant observation are presented as illustrative case material to support the authors view, that ritual procedures are not only defence mechanisms against anxiety, but social acts which generate and convey meaning. Ritual practices described in this analysis include rituals surrounding birth, death, status and power. It is concluded that if nurses wish to change or alter ritual nursing practices in hospital it is necessary to understand their social as well as their psychological meaning.

Anxiety↗
PubMed1981

The 20,000 molecular weight variant of human growth hormone. Preparation and some physical and chemical properties.

Extracts of frozen human pituitary glands contain, in addition to normal growth hormone (hGH (Mr = 22,005), a variant of lower molecular weight (approximately 20,000) (Singh, R. N. P., Seavey, B. K. and Lewis, U. J. (1974) Endocrinol. Res. Commun. 1, 449-464). This material, designated 20K hGH, occurs in various forms with the majority present as a heterologous dimer of Mr = 20,000 hGH and hGH. The purification scheme for obtaining the variant from these fractions is described. Analysis of Mr = 20,000 hGH in the sequenator has confirmed that residues 32-46 of hGH are absent in Mr = 20,000 hGH. The structures of hGH and Mr = 20,000 hGH have been compared by 1H nuclear magnetic resonance spectroscopy, circular dichroism, spectrophotometric titration, and nitration of the tyrosine residues by tetranitromethane. The results indicate that the folding of the polypeptide chain of the two proteins is similar but not identical.

Amino Acid Sequence↗
PubMed1981

The isolation of human pituitary hormones from frozen glands.

An integrated scheme for the extraction and purification of human pituitary hormones from frozen glands has been developed. The method yields 6.3 mg/gland of monomeric pyrogen-free GH in a single fraction, with a potency of 2.5-3.1 IU/mg, as estimated by bioassay, radioreceptor assay, and RIA, with a further 0.6 mg/gland recovered from side fractions. A glycoprotein fraction yields 200 IU FSH, 500 IU LH, and 0.2 IU TSH per gland, as estimated by RIA. Additional hormones purified from side fractions include PRL, lipotropins, and the 20,000 molecular weight variant of human GH.

Chromatography, DEAE-Cellulose↗
PubMed1979

The conformation of histone H5 bound to DNA. Maintenance of the globular structure after binding.

Trypsin digestion is used to investigate the conformation of histone H5 when bound to DNA. A central region of H5 comprising residues (22--100) is found to be resistant to digestion and it is concluded that this region is compacted whilst the remaining N- and C-terminal regions are more extended. Since this is the same result found previously for the free solution conformation of histone H5 it follows that a 3-domain structure is preserved on DNA binding. The binding of H5 and the central region (22--100) to DNA is also studied using proton magnetic resonance (270 MHz) and a precipitation approach. It is concluded that all 3 domains of H5 bind to DNA at low ionic strengths. The central domain (residues 22--100) is released at 0.3--0.4 M NaCl, but 0.7 M NaCl is required to release the N- and C-terminal regions. Comparison is made of H5 binding to DNA with that of the related histone H1.

Animals↗
PubMed1978

The conformation of histone H5. Isolation and characterisation of the globular segment.

Treatment of chicken erythrocyte histone H5 with trypsin in a high-ionic-strength medium results in very rapid initial digestion and the formation of a 'limiting' resistant product peptide. Under these solution conditions the H5 molecule is maximally folded by spectroscopic criteria and it is concluded that the resistant peptide, GH5, represents a globular folded region of the molecule whilst the rapidly digested parts are disordered. The peptide GH5 is shown to comprise the sequence 22-100. In support of this conclusion it is shown that whilst intact histone H5 is hydrodynamically far from being a compact globular shape, peptide GH5 is approximately spherical by hydrodynamic and scattering criteria. Further more, peptide GH5 retains all the alpha-helical structure of intact H5 (circular dichroism) and appears to also maintain all the tertiary structure (nuclear magnetic resonance). It follows that in solution at high ionic strength, histone H5 consists of three domains: an N-terminal disordered region 1-21, a compact globular central domain 22-100 and a long disordered C-terminal chain 101-185. Structural parallels are drawn with the three-domain structure of the histone H1 molecule.

Amino Acid Sequence↗
PubMed1978

A nuclear-magnetic-resonance study of the globular structure of the H5 histone.

The structure of the globular region of the chicken erythrocyte H5 histone has been studied by 270-MHz proton magnetic resonance. The aromatic resonances have been partially assigned by a combination of selective deuteration and iodination with the nuclear magnetic resonance spectroscopy. Detailed titration studies have revealed interactions between residues in the structure. A technique involving the measurement of small nuclear Overhauser effects has enabled the assignment of the aromatic residues causing the perturbation of the ring-current-shifted methyl resonances occurring in the upfield region of the spectrum. Spin-decoupling experiments on these peaks has enabled a partial assignment of shifted methyl resonances. The results support the notion that the histone H5 globular structure is different from that of the homologous histone H1 molecule.

Animals↗
PubMed1977

Studies on the role and mode of operation of the very-lysine-rich histone H1 in eukaryote chromatin. The three structural regions of the histone H1 molecule.

Limited digestion with trypsin of both calf thymus H1 histone and the fragment 1--120 of the H1 molecule has resulted in the isolation of the fragment 35--120. This fragment assumes a globular structure under physiological conditions of pH and ionic strength. The variable N-terminal portion of the molecule, up to residue 34, is not required for the formation of the H1 globular structure. Proton nuclear magnetic resonance (NMR) and ultracentrifugation studies show that the H1 histone molecule consists of three distinct structural domains under structuring conditions: a random coil 'nose' consisting of 35 to 40 residues from the N-terminal end; a globular 'head' involving the next approximately 80 residues; and a random-coil 'tail' of the remainder of the molecule.

Amino Acids↗
PubMed1976

Studies on the role and mode of operation of the very-lysine-rich histone H1 in eukaryote chromatin. The isolation of the globular and non-globular regions of the histone H1 molecule.

Digestion of calf thymus H1 histone with thrombin cleaves the molecule at the sequence -(Pro)-Lys-Lys-Ala-, corresponding to a point approximately 122 residues from the N-terminus (about 56% along the molecule). The N-terminal fragment is shown by proton nuclear magnetic resonance (NMR) to possess the globular structure of the intact histome H1 molecule, whereas the C-terminal fragment appears to possess little or no structure. The N-terminal fragment separates into two peaks on an ion-exchange column, one of which is shown to originate from a single subfraction of calf thymus histone H1 and the other to originate from the other subfractions, by detailed comparison of the NMR spectra. It thus seems that the structure of the H1 histone in solution under physiological conditions consists of a globular head with a highly basic random coil tail. It is suggested that the globular head has a specific binding site on the subunit structure of the chromosome.

Amino Acids↗
PubMed1975

Studies on the role and mode of operation of the very-lysine-rich histone H1 (F1) in eukaryote chromatin. The properties of the N-terminal and C-terminal halves of histone H1.

Restricted chymotrypsin digestion of calf thymus H1 histone gives two fragments, residues 1--106 and 107--C-terminal. These were studied by proton magnetic resonance and circular dichroism. The N-terminal fragment exhibited some salt-induced structure in aqueous solution, but this did not parallel the globular structure of the intact H1 molecule. Comparison of circular dichroism results with helix predictions for this portion of the molecule suggests that the secondary structure may be the same in this fragment as it is in the corresponding region of the whole molecule. The C-terminal fragments show very little salt-induced structure. The N-terminal fragments binds to DNA very weakly, but the C-terminal fragment binds as strongly as the whole molecule. In the C-terminal fragment, about one quarter of the lysine residues are not bound to the DNA in water, but initial increase of salt concentration causes them to become bound. This increasing binding occurs under the same ionic conditions that cause chromatin condensation and condensation of H1 - DNA complexes, and it is suggested that there may be a connection between these phenomena.

Amino Acids↗
PubMed1975

Studies on the role and mode of operation of the very-lysine-rich histone H1 (F1) in eukaryote chromatin. The conformation of histone H1.

Proton magnetic resonance, circular dichroism and other studies of whole and cleaved calf thymus histone H1 (formerly F1) reveal the presence of specific folded structures in the region approximately from residue 40--115. Ionic, hydrogen-bond and hydrophobic interactions all appear to contribute to the stability of the structure, which is predicted to contain alpha-helices in regions 42--55 and 58--75. No evidence was found for beta-structures, either inter or intramolecular, or for any structure formation outside the region 40--115. At 18 degrees C and a protein concentration of 2 mM the first-order exchange rate between random-coil and structured forms is slower than 80 s-1; at 40 degrees C the exchange rate is faster than 330 s-1.

Animals↗