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H D Niall

Publications and source records attributed to H D Niall.

At least 37 records · Page 2Linked to original sources

The evolution of peptide hormones.

Despite limitations in our present knowledge it is already possible to discern the main features of peptide hormone evolution, since the same mechanisms (and indeed the same hormone molecules) function in many different ways. This underlying unity of organization has its basis in the tendency of biochemical networks, once established, to survive and diversify. The most surprising recent findings in endocrinology have been the discovery of vertebrate peptide hormones in multiple sites within the same organism, and the reports, persuasive but requiring confirmation, of vertebrate hormones in primitive unicellular organisms (20, 20a). Perhaps the major challenge for the future is to define the roles and interactions of the many peptide hormones identified in brain (18). The most primitive bacteria and the human brain, though an enormous evolutionary distance apart, may have more in common than we have recognized until now. As Axelrod & Hamilton have pointed out in a recent provocative article, "The Evolution of Cooperation" (1), bacteria, though lacking a brain, are capable of adaptive behavior that can be analysed in terms of game theory. It is clear that we can learn a great deal about the whole evolutionary process from a study of the versatile and durable peptide hormones molecules.

Animals↗

Amino acid sequence of p15 from avian myeloblastosis virus complex.

The complete amino acid sequence of the p15 gag protein from avian myeloblastosis virus (AMV) complex has been determined by sequential Edman degradation of the intact molecule and of peptide fragments generated by limited tryptic cleavage, cleavage with staphylococcal protease, and cyanogen bromide cleavage. AMV p15 is a single-chain protein containing 124 amino acids. The charged amino acids tend to be clustered in the primary structure. p15 contains a single cysteine at position 113 which may be essential for the p15 associated proteolytic activity. However, p15 shows no appreciable sequence homology with papain or other classical thiol proteases.

Amino Acid Sequence↗

Limited sequence homology between porcine and rat relaxins: implications for physiological studies.

We have isolated rat relaxin as a single chromatographic component and determined its complete amino acid sequence. Comparison with porcine relaxin reveals a clear sequence homology but the extent of this is much more limited than expected, with only 21 of 53 residues (39.6%) in corresponding positions being identical. In contrast 47 of 51 residues (92.2%) are identical between porcine and rat insulins. These findings suggest that relaxin has been less constrained to conserve its structural features during evolution than has insulin, since the original duplication of a common ancestral gene. Relaxins as a group (and rat and porcine relaxins in particular) show poor immunological cross-reactivity despite a common biological activity. The finding of extensive sequence differences as well as conserved residues between porcine and rat relaxins provides a possible structural basis for these observations, and should facilitate identification of a localized receptor-binding surface region of relaxin. A further important implication of this limited sequence homology is that physiological (and clinical) studies with relaxins should whenever possible be carried out with hormone preparations from the same species.

Amino Acid Sequence↗

Use of an octadecylsilica purification method minimizes proteolysis during isolation of porcine and rat relaxins.

Previous attempts to purify relaxin from pregnant sow and rat ovaries have led to the identification of multiple components with very similar molecular weights (around 6000 daltons) and similar, but not identical amino acid compositions. It has been unclear whether these multiple forms of relaxin represented different allelic forms (isohormones), intermediates in a prohormone-hormone conversion, different hormones in their own right responsible for effects on different target tissues, or degradation products. We have applied a purification system using adsorption to small octadecylsilica (ODS) columns to reduce drastically the degree of heterogeneity. Both pig and rat relaxins are isolated as single major components with defined amino acid sequences and with minimal contamination by other molecular forms. These findings indicate that the relaxin heterogeneity previously observed is due to proteolysis of a single major stored form during conventional isolation procedures.

Animals↗

Evidence for proteolysis during purification of relaxin from pregnant sow ovaries.

The proteolytic degradation of relaxin during its isolation from pregnant sow ovaries has been examined. Ovaries from pregnant sows were selected and divided into three groups according to the stages of pregnancy. Each group was extracted with and without protease inhibitors. It was found that protease(s) were present in all groups of ovaries and that a 2-4 fold increase in yield of total relaxin was obtained when isolation and purification was carried out in the presence of protease inhibitors. However the ratio of the three forms of relaxin remain unchanged.

Animals↗

Relaxin: an insulin-related growth factor.

Relaxin is an ovarian peptide that is released just prior to parturition to effect changes in the tissues of the birth canal that aid in the delivery of the fetus. Structural studies established that the two constituent polypeptide chains, composed of 22 and 30 amino acids, are joined by two interchain disulfide bonds with an additional third intrachain bridge. As well as the identical pattern of disulfide bonds, relaxin shows an overall 25% identity with insulin. Furthermore, the sequence of relaxin can be incorporated into the known three-dimensional structure of insulin without significant distortion of the main polypeptide chain backbone. The discovery of the insulin-relatedness of relaxin brings to three the number of growth factors that share a common structural gene precursor with insulin. Nerve growth factor and insulin-like growth factor have already been so identified. Inclusion in this hormone family suggests that the mechanism of action may involve internalization as well as complexation with cell surface receptors of target cells.

Amino Acid Sequence↗

Synthesis, purification, and chemical characterization of the amino-terminal 1-34 fragment of bovine parathyroid hormone synthesized by the solid-phase procedure.

Peptides prepared by solid-phase synthesis during a systematic study of structure-activity relations in parathyroid hormone have been subjected, after sequential purifications, to rigorous analysis of purity by a variety of analytical techniques including sequence analysis by the Edman procedure. The present paper undertakes a critical appraisal of the utility of different tests of peptide purity and the use of the procedures in guiding and monitoring optimal synthesis strategies. Sequence analysis of a peptide representing the amino-terminal 34 residues of bovine parathyroid hormone (bPTH-(1-34)) revealed the presence of at least 30% of contaminating error peptides which were undetected by other analytical procedures. The major contaminant was identified as a peptid in which glutamine at position 29 was deleted. A repeat synthesis using fluorescamine rather than ninhydrin to monitor the coupling reaction resulted in a preparation that lacked the contaminant resulting from deletion. These findings demonstrate the particular value of sequence analysis in the evaluation of purity of peptides synthesized by the solid-phase technique.

Adenylyl Cyclases↗

Purification, characterization, and partial amino acid sequence of nerve growth factor from cobra venom.

The nerve growth factor (NGF) from Naja naja (cobra) venom has been purified and its structure compared to the NGF from mouse submaxillary gland. A two-step purification procedure has been devised, consisting of a gel filtration step in 1 M acetic acid followed by chromatography of the active pool on carboxymethylcellulose at pH 5. The molecular weight of the native protein was found to be 28000, and this value was reduced by approximately one-half under denaturing conditions. These values are comparable to those obtained for mouse 2.5S or betaNGF. Tryptic peptide maps of S-[14C]carboxymethyl NGF gave the number of labeled peptides expected for a structure composed of two identical or very similar subunits. Thus, the quaternary structures of mouse and cobra NGF are the same. Cyanogen bromide (CNBr) treatment of Naja naja NGF produced three fragments, of which two were purified to homogeneity. These fragments and the whole protein were analyzed in the automated protein Sequencer. The amino-terminal CNBr fragment of the protein was also subjected to digestion by thermolysin and the resultant peptides were purified and characterized. These data plus those from the characterization of the tryptic peptides provided the basis of the construction of a tentative primary structure of Naja naja NGF which is approximately 60% identical with mouse NGF.

Amino Acid Sequence↗

Angiotensin II analogues and aldosterone secretion in sodium-deficient sheep.

1. The angiotensin analogues Sar1-Ala8-angiotensin II (AII), Sar1-Ile8-AII, Sar1-Leu8-AII, Sar1-Thr8-AII, [Des1-Asp]-Ile8-AII and [Des1-Asp]-Sar2-Ile8-AII and converting enzyme inhibitor (SQ 80221) infused by intra-adrenal arterial infusion had no effect on aldosterone secretion in sodium-deficient sheep at doses in excess of those shown to block exogenous angiotensin II or III infusion. 2. It is suggested that the intrinsic agonist activity of the analogues may fulfil the requirements for a permissive role for angiotensin in the aldosterone response to sodium deficiency.

Aldosterone↗

Effect of adrenal arterial infusion of saralasin (P113) on aldosterone secretion.

To test the role of the renin/angiotensin system in aldosterone regulation, Sar1-Ala8-angiotensin II (P113) was infused into the arterial blood supply of transplanted adrenal glands in conscious sheep. Effects on the aldosterone response to infused angiotensin II and III in sodium replete sheep were compared with effects in sodium deficiency. Adrenal arterial infusion of P113 up to 1,000 mug/h for 1-2h did not consistently alter the high aldosterone secretion rates of sodium-deficient sheep. However, infusion of P113 at 10 mug/h or more abolished aldosterone responses to angiotensin II infusion that caused high physiological blood levels of angiotensin II. These results are against the proposal of a primary proportional causal relationship between blood angiotensin II concentration and aldosterone secretion rate in sodium deficiency. They are also against the possibility that such a primary role is played by blood-borne angiotensin III.

Adrenal Glands↗

High sensitivity automated sequence determination of polypeptides.

We report the development of a high sensitivity Edman method for use in the automated protein sequenator. With the use of a radioactive coupling reagent, [35S]phenylisothiocyanate, and minor modifications in the sequenator program, sequence analyses have been performed on nanomole quantities of protein. The radioactive phenylthiohydantoin derivatives produced in the degradation are identified at the 10 to 100 pmol level by two-dimensional thin layer chromatographic procedures with autoradiography and are quantitated by scintillation counting. This high sensitivity approach, which is about 100 times more sensitive than conventional automated Edman procedures, has allowed continuous amino acid assignments for 15 or more cycles on quantities of protein less than 5 nmol. It has been employed in the NH2-terminal sequence analysis of several proteins whose sequences were previously undetermined.

Amino Acid Sequence↗

The complete amino acid sequence of ubiquitin, an adenylate cyclase stimulating polypeptide probably universal in living cells.

The complete amino acid sequence was determined for bovine ubiquitin, and adenylate cyclase stimulating polypeptide, which is probably represented universally in living cells. Ubiquitin has a molecular weight of 8451 and consists of a single polypeptide chain containing 74 amino acid residues. It contains four arginine residues but no cysteine or trytophan residues. The first 61 amino acid residues were obtained by automated Edman degradations. Tryptic digestion of maleated ubiquitin yielded four peptide fragments that were resolved by molecular sieve chromatography and coded in order of decreasing chain length (MT-1, MT-2, MT-3, and MT-4). The automated sequenator determinations on native ubiquintin provided overlapping sequence data for three of these fragments that gave an order of MT-1, MT-3, and then MT-2; Peptide MT-4, a dipeptide, was therefore assigned to the C terminus, and the placement of peptide MT-2 was corroborated by analysis of data from carboxypeptidase digestions of maleated ubiquitin. Peptide MT-2 was domaleated and sequenced by manual Edman degradations through a single lysine residue. It was cleaved at this residue with trypsin, and the two resultant peptides were separated by ion-exchange chromatography. Manual sequencing of the C-terminal demaleated tryptic peptide of MT-2 completed the sequence of MT-2 and that of native ubiquitin. The sequence of ubiquitin was further confirmed and supported by amino acid and parital sequence anlysis of fragments obtained by digestion of maleated ubiquitin with chymotrypsin or staphylococcal protease.

Adenylyl Cyclases↗

A reinvestigation of the amino-terminal sequence of human parathyroid hormone.

The sequence of the amino-terminal portion of human parathyroid hormone, particularly the identity of residues 22, 28, and 30 (the subject of discrepancies in recent published reports), has been reexamined by two basic methods of structural analysis. A fresh lot of human parathyroid hormone isolated from pooled adenoma tissue was analyzed by Edman degradation with identification of critical residues by thin-layer chromatography and gas-liquid chromatography. In the second approach, -14C or tritiated amino acids were incorporated during biosynthesis of the human hormone in slices of parathyroid glands in vitro; the appropriate amino acid residues were then determined as the -14C or tritiated phenythiohydantoin derivatives of the amino acid after Edman degradation, or by peptide isolation after appropriate cleavage with endopeptidase, or both. The results confirm our previous findings that residue 22 is glutamic acid, residue 28 is leucine, and residue 30 is aspartic acid.

Adenoma↗

Isolation of a polypeptide that has lymphocyte-differentiating properties and is probably represented universally in living cells.

A polypeptide of 8500 molecular weight is described that induces the differentiation of T (thymus-derived) cell and B (bone-marrow-derived) cell immunocytes in vitro, apparently via beta-adrenergic receptors and adenylate cyclase activation. This polypeptide shows a high degree of evolutionary conservation, exhibiting close structural, functional, and immunological similarity when isolated from such diverse origins as cells of mammals and higher plants. This polypeptide was detected in animal cells, yeast, bacteria, and higher plants, and so may well be a universal constituent of living cells.

Adenylyl Cyclases↗