Glenn Hammonds - Academia.edu (original) (raw)

Papers by Glenn Hammonds

Proceedings of The National Academy of Sciences, 1981

The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (be... more The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (beta h-EP). The affinity and number of sites are 1.1 nM and 280,000 per cell, respectively, beta h-EP binding is not inhibited by [Leu]enkephalin or morphine at concentrations up to 0.1 mM; beta h-EP-(6--31) is a potent inhibitor of binding, and camel beta-EP is much less potent. The data suggest the importance of the nonenkephalin segment of the beta h-EP molecule for interaction with the binding site in N18TG2 cells.

International Journal of Peptide and Protein Research, 2009

Opioid receptor selectivity of several β-endorphin (β-EP) analogs which antagonize β-EP-induced a... more Opioid receptor selectivity of several β-endorphin (β-EP) analogs which antagonize β-EP-induced analgesia has been assessed using partially selective binding assays. Although the apparent affinity dissociation constant of β-EP in these assays varies from 0.2 to 360 nm, the potency of β-EP antagonists relative to β-EP remains largely unchanged. It is unlikely that differences in receptor affinities can account for the antagonist properties of these analogs in vivo.

Proceedings of The National Academy of Sciences, 1982

Rat brain membranes (BBM) bind (3-endorphin with high affinity and specificity. We report herein ... more Rat brain membranes (BBM) bind (3-endorphin with high affinity and specificity. We report herein the identification of a high molecular weight ,-endorphin complex (receptor) in extracts of RBM preincubated with tritiated -endorphin, by using the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate (CHAPS). Upon isoelectric focusing, this complex gives a single peak with an isoelectric point (+SEM) of 4.50 ± 0.06. Sucrose density gradient experiments in H20 and 2H20 yield effective partial specific volume (v* = 0.814 cm3/g and sedimentation constant 820,w = 15.6 S. Gel filtration

Science, 1987

Humoral hypercalcemia of malignancy is a common complication of lung and certain other cancers. T... more Humoral hypercalcemia of malignancy is a common complication of lung and certain other cancers. The hypercalcemia results from the actions of tumor factors on bone and kidney. We report here the isolation of full-length complementary DNA clones of a putative hypercalcemia factor, and the expression from the cloned DNA of the active protein in mammalian cells. The clones encode a prepro peptide of 36 amino acids and a mature protein of 141 amino acids that has significant homology with parathyroid hormone in the amino-terminal region. This previously unrecognized hormone may be important in normal as well as abnormal calcium metabolism.

Molecular Endocrinology, 1991

The human osteoinductive proteins BMP-2a and BMP-2b have been cloned and expressed in mammalian c... more The human osteoinductive proteins BMP-2a and BMP-2b have been cloned and expressed in mammalian cells. In order to improve expression levels we examined the role of the proregion in assembly and export. Use of the BMP-2a proregion combined with the mature region of BMP-2b leads to dramatically improved expression of mature BMP-2b. Mature BMP-2b has been purified to near homogeneity from the BMP-2a/2b hybrid, and its structural properties and biological activity determined. Recombinant mature BMP-2b homodimer elicits bone formation in vivo.

Biopolymers, 1976

The existence and distribution of vertical, interacting edges on cylindrical surfaces, which were... more The existence and distribution of vertical, interacting edges on cylindrical surfaces, which were derived from the basic structural features of the collagen molecule, have been examined as a function of the angle θ of the superhelix. Over the range θ = 26°–33° and near θ = 36°, the strongest intermolecular association was detected at 29.95°. This model exhibited three similar, self-interacting edges, separated by approximately 120°. The potential for three similar but nonidentical edges lies in the geometry of the collagen molecule, but the existence of such edges and their ability to self-associate is dependent upon the unique amino acid sequence. The relative strengths of association of each edge with itself and with the other edges are not identical, but the present analysis does not permit a judgment as to whether this variation could result in functional specificity in such interactions. All three chains participate in each edge though their contribution to the relative interaction strength is not identical.These edges showed distinct maxima when displaced past one another by integral intervals of D, where D = 233 residue positions (quarter-stagger). The strongest relative interaction occurs at a displacement of 1D.

Journal of Cellular Physiology, 1993

The effect of transforming growth factor-pl (TGFP1) on three developmental stages of megakaryocyt... more The effect of transforming growth factor-pl (TGFP1) on three developmental stages of megakaryocytopoiesis was investigated. Using a murine bone marrow agar culture system, titrated doses of TGFPl were added to cultures assaying primitive high pro1 iferative megakaryocyte progenitors, committed megakaryocyte precursors, and nondividing, endoreduplicating megakaryocytes. The growth of high proliferative megakaryocyte colony-forming cells (HPP-CFU-Mk) that require the growth factors interleukins-I, 3 and 6 (IL-1 + IL-3 + IL-6) for

Analytical Biochemistry, 1981

... ANA[ YFICAI-FSIOCHFMISTRY 114, 75-84 (1981) Interaction of Tritiated f3-Endorphin with Rat Br... more ... ANA[ YFICAI-FSIOCHFMISTRY 114, 75-84 (1981) Interaction of Tritiated f3-Endorphin with Rat Brain Membranes R. GLENN HAMMONDS, JR.,*'I NICHOLAS LING,t AND DAVID PUETT*" *Department of Biochemistry ... (B) Kinetics of dissociation of F3-[MeTI ]endorphin (0.5 nM ...

Journal of Bone and Mineral Research, 1994

In many cell systems, cell-cell and cell-matrix interactions are mediated by integrins, a family ... more In many cell systems, cell-cell and cell-matrix interactions are mediated by integrins, a family of cell surface heterodimeric glycoprotein receptors. Osteoclast integrins may play a role in the process of bone resorption. Osteoclasts express the αv and β3 subunits of the vitronectin receptor and adhere to a wide range of proteins in vitro, all of which contain the amino acid sequence Arg-Gly-Asp (RGD), an adhesion site recognition sequence common to many protein ligands that bind to integrins. The effect of kistrin, an RGD-containing snake venom protein, on osteoclast-mediated bone resorption was investigated in vivo and in vitro. When kistrin was infused into normocalcemic and hypercalcemic mice, serum calcium was significantly lowered at 3 and 6 h after the start of infusion, indicating an inhibitory effect on osteoclast activity in vivo. In vitro, kistrin potently inhibited bone resorption by isolated rat osteoclasts cultured on slices of bovine bone, and kistrin also inhibited the attachment of 293 cells expressing recombinant human αvβ3 to fibrinogen (IC50 = 1 nM). These results indicate the potential therapeutic use of RGD-containing molecules for hypercalcemia of malignancy or for other disorders associated with bone loss.

International Journal of Peptide and Protein Research, 2009

βh-Endorphin-(1–27) (I), [Ac-Tyr1]-βh-endorphin-(1–27) (II), [Gln8]-βh-endor-phin-(1–27) (III), a... more βh-Endorphin-(1–27) (I), [Ac-Tyr1]-βh-endorphin-(1–27) (II), [Gln8]-βh-endor-phin-(1–27) (III), and [Ac-Tyr1, Gln8]-βh-endorphin-(1–27) (IV) were synthe sized by the solid-phase method. The binding potency of I-IV to rat brain membrane preparations was measured by radioreceptor binding assay. The relative potencies were: βh-endorphin, 100; I, 30; II, 0.04; III, 90; IV, 0.07.

European Journal of Biochemistry, 1977

It is shown that the method proposed by Baker and Isenberg [Biochemistry, 15, 629 (1976)l for est... more It is shown that the method proposed by Baker and Isenberg [Biochemistry, 15, 629 (1976)l for estimating secondary structure composition of proteins from circular dichroic spectra is a leastsquares fitting technique. Estimates obtained by this method for myoglobin, lysozyme, lactate dehydrogenase, papain, and ribonuclease are not substantively different from those obtained using unconstrained linear least squares.

[

Endocrinology, 1991

We have recently shown that a carboxyl-terminal sequence of parathyroid hormone-related protein, ... more We have recently shown that a carboxyl-terminal sequence of parathyroid hormone-related protein, PTHrP[107-139], is a potent direct inhibitor of osteoclastic bone resorption. We now report that this bone resorption inhibitory activity, which we have called osteostatin, is entirely contained within the highly conserved pentapeptide PTHrP[107-111]. Our results indicate that processing at residue 106 and a free amino terminus is required for full activity of the peptide. The retroinverted peptide is considerably less potent than the parent peptide. The retention of full biological activity in such a short fragment was unexpected. This data provides the basis for the development of further analogs with potential therapeutic use in disorders associated with increased osteoclastic bone resorption.

Proceedings of The National Academy of Sciences, 1984

beta h-Endorphin-(1-27), a naturally occurring fragment of human beta-endorphin (beta h-endorphin... more beta h-Endorphin-(1-27), a naturally occurring fragment of human beta-endorphin (beta h-endorphin), diminishes the analgesic effect of beta h-endorphin when coinjected intra-cerebroventricularly into mice. A parallel shift in the dose-response curve of beta h-endorphin in the presence of beta h-endorphin-(1-27) suggests competition at the same site. The potency of beta h-endorphin-(1-27) in antagonizing analgesia is greater than 4 times greater than that of the opiate antagonist naloxone.

International Journal of Peptide and Protein Research, 2009

Four analogs of human β-endorphin (βh-EP) were synthesized by the solid-phase method: βh-EP-(1–17... more Four analogs of human β-endorphin (βh-EP) were synthesized by the solid-phase method: βh-EP-(1–17) (I), [D-Ala2]-βh-EP-(1–17) (II), [Gln8]-βh-EP-(1–17) (III) and [D-Ala2, Gln8]-βh-EP-(1–17) (IV). Measurement in a radio-receptor binding assay with use of tritiated βh-EP as primary ligand gave relative potencies as follows: Met-enkephalin, 100; I, 33; II, 47; III, 889; IV, 123; βh-endorphin, 2253.

International Journal of Peptide and Protein Research, 2009

The potency of a series of synthetic analogs of β-endorphin in inhibiting binding of [3H2-Tyr27]-... more The potency of a series of synthetic analogs of β-endorphin in inhibiting binding of [3H2-Tyr27]-βh-endorphin to either rat brain membranes or neuroblastoma x glioma hybrid cells (NG108–15) has been determined and compared with the previously determined analgesic potency. There is a very good correlation between inhibitory potency in membranes and cells, but the correlation between analgesic potency and inhibitory potency in either membranes or cells is not as good.

International Journal of Peptide and Protein Research, 2009

A double-headed analog of human β-endorphin (βh-EP), N, N'-bis (β-endorphinyl)-cystine (II), has ... more A double-headed analog of human β-endorphin (βh-EP), N, N'-bis (β-endorphinyl)-cystine (II), has been synthesized by the solid-phase method, along with βh-EP-Cys(CH2CONH2)-OH (I) and (Tyr31]-βh-EP (III). Their relative potencies in a radioreceptor-binding assay were: Bh-EP, 100; II, 235; I, 170; and III, 204. In the tail-flick test for analgesic activity their relative potencies were: βhEP, 100; II, 86; I, 93; and III, 116.

International Journal of Peptide and Protein Research, 2009

Analgesic potencies of turkey, ostrich and des-acetyl salmon β-endorphins have been measured in t... more Analgesic potencies of turkey, ostrich and des-acetyl salmon β-endorphins have been measured in the tail-flick test and binding affinities determined by radioreceptor assay. The duration of analgesia and the slope of the dose-response curves generated by these peptides are similar to those elicited by mammalian β-endorphins. This suggests that they act in vivo and in vitro on the same population of opiate receptors. The ratio of binding to analgesic potencies observed for these peptides varies nearly sixfold. Structure-activity analysis suggests that a basic side-chain at position 9 is required in order to produce a high opiate activity both in vivo and in vitro. A reexamination of the biological activities of camel β-endorphin shows that the analgesic potency and binding affinity of this peptide are respectively 1.7 and 2.7 times higher than human β-endorphin. His-27 and/or Gln-31 may contribute to this increased potency. The dissociation of radioreceptor binding affinity from analgesic potency in these naturally occurring β-endorphin homologs suggests that either the conditions under which the binding assay is performed mask the true binding potency in the brain or that, once bound to the appropriate receptor, these homologs do not possess equal ability to produce biological effects.

Proceedings of The National Academy of Sciences, 1981

The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (be... more The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (beta h-EP). The affinity and number of sites are 1.1 nM and 280,000 per cell, respectively, beta h-EP binding is not inhibited by [Leu]enkephalin or morphine at concentrations up to 0.1 mM; beta h-EP-(6--31) is a potent inhibitor of binding, and camel beta-EP is much less potent. The data suggest the importance of the nonenkephalin segment of the beta h-EP molecule for interaction with the binding site in N18TG2 cells.

International Journal of Peptide and Protein Research, 2009

Opioid receptor selectivity of several β-endorphin (β-EP) analogs which antagonize β-EP-induced a... more Opioid receptor selectivity of several β-endorphin (β-EP) analogs which antagonize β-EP-induced analgesia has been assessed using partially selective binding assays. Although the apparent affinity dissociation constant of β-EP in these assays varies from 0.2 to 360 nm, the potency of β-EP antagonists relative to β-EP remains largely unchanged. It is unlikely that differences in receptor affinities can account for the antagonist properties of these analogs in vivo.

Proceedings of The National Academy of Sciences, 1982

Rat brain membranes (BBM) bind (3-endorphin with high affinity and specificity. We report herein ... more Rat brain membranes (BBM) bind (3-endorphin with high affinity and specificity. We report herein the identification of a high molecular weight ,-endorphin complex (receptor) in extracts of RBM preincubated with tritiated -endorphin, by using the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate (CHAPS). Upon isoelectric focusing, this complex gives a single peak with an isoelectric point (+SEM) of 4.50 ± 0.06. Sucrose density gradient experiments in H20 and 2H20 yield effective partial specific volume (v* = 0.814 cm3/g and sedimentation constant 820,w = 15.6 S. Gel filtration

Science, 1987

Humoral hypercalcemia of malignancy is a common complication of lung and certain other cancers. T... more Humoral hypercalcemia of malignancy is a common complication of lung and certain other cancers. The hypercalcemia results from the actions of tumor factors on bone and kidney. We report here the isolation of full-length complementary DNA clones of a putative hypercalcemia factor, and the expression from the cloned DNA of the active protein in mammalian cells. The clones encode a prepro peptide of 36 amino acids and a mature protein of 141 amino acids that has significant homology with parathyroid hormone in the amino-terminal region. This previously unrecognized hormone may be important in normal as well as abnormal calcium metabolism.

Molecular Endocrinology, 1991

The human osteoinductive proteins BMP-2a and BMP-2b have been cloned and expressed in mammalian c... more The human osteoinductive proteins BMP-2a and BMP-2b have been cloned and expressed in mammalian cells. In order to improve expression levels we examined the role of the proregion in assembly and export. Use of the BMP-2a proregion combined with the mature region of BMP-2b leads to dramatically improved expression of mature BMP-2b. Mature BMP-2b has been purified to near homogeneity from the BMP-2a/2b hybrid, and its structural properties and biological activity determined. Recombinant mature BMP-2b homodimer elicits bone formation in vivo.

Biopolymers, 1976

The existence and distribution of vertical, interacting edges on cylindrical surfaces, which were... more The existence and distribution of vertical, interacting edges on cylindrical surfaces, which were derived from the basic structural features of the collagen molecule, have been examined as a function of the angle θ of the superhelix. Over the range θ = 26°–33° and near θ = 36°, the strongest intermolecular association was detected at 29.95°. This model exhibited three similar, self-interacting edges, separated by approximately 120°. The potential for three similar but nonidentical edges lies in the geometry of the collagen molecule, but the existence of such edges and their ability to self-associate is dependent upon the unique amino acid sequence. The relative strengths of association of each edge with itself and with the other edges are not identical, but the present analysis does not permit a judgment as to whether this variation could result in functional specificity in such interactions. All three chains participate in each edge though their contribution to the relative interaction strength is not identical.These edges showed distinct maxima when displaced past one another by integral intervals of D, where D = 233 residue positions (quarter-stagger). The strongest relative interaction occurs at a displacement of 1D.

Journal of Cellular Physiology, 1993

The effect of transforming growth factor-pl (TGFP1) on three developmental stages of megakaryocyt... more The effect of transforming growth factor-pl (TGFP1) on three developmental stages of megakaryocytopoiesis was investigated. Using a murine bone marrow agar culture system, titrated doses of TGFPl were added to cultures assaying primitive high pro1 iferative megakaryocyte progenitors, committed megakaryocyte precursors, and nondividing, endoreduplicating megakaryocytes. The growth of high proliferative megakaryocyte colony-forming cells (HPP-CFU-Mk) that require the growth factors interleukins-I, 3 and 6 (IL-1 + IL-3 + IL-6) for

Analytical Biochemistry, 1981

... ANA[ YFICAI-FSIOCHFMISTRY 114, 75-84 (1981) Interaction of Tritiated f3-Endorphin with Rat Br... more ... ANA[ YFICAI-FSIOCHFMISTRY 114, 75-84 (1981) Interaction of Tritiated f3-Endorphin with Rat Brain Membranes R. GLENN HAMMONDS, JR.,*'I NICHOLAS LING,t AND DAVID PUETT*" *Department of Biochemistry ... (B) Kinetics of dissociation of F3-[MeTI ]endorphin (0.5 nM ...

Journal of Bone and Mineral Research, 1994

In many cell systems, cell-cell and cell-matrix interactions are mediated by integrins, a family ... more In many cell systems, cell-cell and cell-matrix interactions are mediated by integrins, a family of cell surface heterodimeric glycoprotein receptors. Osteoclast integrins may play a role in the process of bone resorption. Osteoclasts express the αv and β3 subunits of the vitronectin receptor and adhere to a wide range of proteins in vitro, all of which contain the amino acid sequence Arg-Gly-Asp (RGD), an adhesion site recognition sequence common to many protein ligands that bind to integrins. The effect of kistrin, an RGD-containing snake venom protein, on osteoclast-mediated bone resorption was investigated in vivo and in vitro. When kistrin was infused into normocalcemic and hypercalcemic mice, serum calcium was significantly lowered at 3 and 6 h after the start of infusion, indicating an inhibitory effect on osteoclast activity in vivo. In vitro, kistrin potently inhibited bone resorption by isolated rat osteoclasts cultured on slices of bovine bone, and kistrin also inhibited the attachment of 293 cells expressing recombinant human αvβ3 to fibrinogen (IC50 = 1 nM). These results indicate the potential therapeutic use of RGD-containing molecules for hypercalcemia of malignancy or for other disorders associated with bone loss.

International Journal of Peptide and Protein Research, 2009

βh-Endorphin-(1–27) (I), [Ac-Tyr1]-βh-endorphin-(1–27) (II), [Gln8]-βh-endor-phin-(1–27) (III), a... more βh-Endorphin-(1–27) (I), [Ac-Tyr1]-βh-endorphin-(1–27) (II), [Gln8]-βh-endor-phin-(1–27) (III), and [Ac-Tyr1, Gln8]-βh-endorphin-(1–27) (IV) were synthe sized by the solid-phase method. The binding potency of I-IV to rat brain membrane preparations was measured by radioreceptor binding assay. The relative potencies were: βh-endorphin, 100; I, 30; II, 0.04; III, 90; IV, 0.07.

European Journal of Biochemistry, 1977

It is shown that the method proposed by Baker and Isenberg [Biochemistry, 15, 629 (1976)l for est... more It is shown that the method proposed by Baker and Isenberg [Biochemistry, 15, 629 (1976)l for estimating secondary structure composition of proteins from circular dichroic spectra is a leastsquares fitting technique. Estimates obtained by this method for myoglobin, lysozyme, lactate dehydrogenase, papain, and ribonuclease are not substantively different from those obtained using unconstrained linear least squares.

[

Endocrinology, 1991

We have recently shown that a carboxyl-terminal sequence of parathyroid hormone-related protein, ... more We have recently shown that a carboxyl-terminal sequence of parathyroid hormone-related protein, PTHrP[107-139], is a potent direct inhibitor of osteoclastic bone resorption. We now report that this bone resorption inhibitory activity, which we have called osteostatin, is entirely contained within the highly conserved pentapeptide PTHrP[107-111]. Our results indicate that processing at residue 106 and a free amino terminus is required for full activity of the peptide. The retroinverted peptide is considerably less potent than the parent peptide. The retention of full biological activity in such a short fragment was unexpected. This data provides the basis for the development of further analogs with potential therapeutic use in disorders associated with increased osteoclastic bone resorption.

Proceedings of The National Academy of Sciences, 1984

beta h-Endorphin-(1-27), a naturally occurring fragment of human beta-endorphin (beta h-endorphin... more beta h-Endorphin-(1-27), a naturally occurring fragment of human beta-endorphin (beta h-endorphin), diminishes the analgesic effect of beta h-endorphin when coinjected intra-cerebroventricularly into mice. A parallel shift in the dose-response curve of beta h-endorphin in the presence of beta h-endorphin-(1-27) suggests competition at the same site. The potency of beta h-endorphin-(1-27) in antagonizing analgesia is greater than 4 times greater than that of the opiate antagonist naloxone.

International Journal of Peptide and Protein Research, 2009

Four analogs of human β-endorphin (βh-EP) were synthesized by the solid-phase method: βh-EP-(1–17... more Four analogs of human β-endorphin (βh-EP) were synthesized by the solid-phase method: βh-EP-(1–17) (I), [D-Ala2]-βh-EP-(1–17) (II), [Gln8]-βh-EP-(1–17) (III) and [D-Ala2, Gln8]-βh-EP-(1–17) (IV). Measurement in a radio-receptor binding assay with use of tritiated βh-EP as primary ligand gave relative potencies as follows: Met-enkephalin, 100; I, 33; II, 47; III, 889; IV, 123; βh-endorphin, 2253.

International Journal of Peptide and Protein Research, 2009

The potency of a series of synthetic analogs of β-endorphin in inhibiting binding of [3H2-Tyr27]-... more The potency of a series of synthetic analogs of β-endorphin in inhibiting binding of [3H2-Tyr27]-βh-endorphin to either rat brain membranes or neuroblastoma x glioma hybrid cells (NG108–15) has been determined and compared with the previously determined analgesic potency. There is a very good correlation between inhibitory potency in membranes and cells, but the correlation between analgesic potency and inhibitory potency in either membranes or cells is not as good.

International Journal of Peptide and Protein Research, 2009

A double-headed analog of human β-endorphin (βh-EP), N, N'-bis (β-endorphinyl)-cystine (II), has ... more A double-headed analog of human β-endorphin (βh-EP), N, N'-bis (β-endorphinyl)-cystine (II), has been synthesized by the solid-phase method, along with βh-EP-Cys(CH2CONH2)-OH (I) and (Tyr31]-βh-EP (III). Their relative potencies in a radioreceptor-binding assay were: Bh-EP, 100; II, 235; I, 170; and III, 204. In the tail-flick test for analgesic activity their relative potencies were: βhEP, 100; II, 86; I, 93; and III, 116.

International Journal of Peptide and Protein Research, 2009

Analgesic potencies of turkey, ostrich and des-acetyl salmon β-endorphins have been measured in t... more Analgesic potencies of turkey, ostrich and des-acetyl salmon β-endorphins have been measured in the tail-flick test and binding affinities determined by radioreceptor assay. The duration of analgesia and the slope of the dose-response curves generated by these peptides are similar to those elicited by mammalian β-endorphins. This suggests that they act in vivo and in vitro on the same population of opiate receptors. The ratio of binding to analgesic potencies observed for these peptides varies nearly sixfold. Structure-activity analysis suggests that a basic side-chain at position 9 is required in order to produce a high opiate activity both in vivo and in vitro. A reexamination of the biological activities of camel β-endorphin shows that the analgesic potency and binding affinity of this peptide are respectively 1.7 and 2.7 times higher than human β-endorphin. His-27 and/or Gln-31 may contribute to this increased potency. The dissociation of radioreceptor binding affinity from analgesic potency in these naturally occurring β-endorphin homologs suggests that either the conditions under which the binding assay is performed mask the true binding potency in the brain or that, once bound to the appropriate receptor, these homologs do not possess equal ability to produce biological effects.