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M D Hollenberg

Publications and source records attributed to M D Hollenberg.

210 records · Page 12Linked to original sources
PubMed1969

Isolation of a third bovine neurophysin.

1. A third native hormone-binding protein, neurophysin-C, has been isolated from acetone-desiccated bovine pituitary posterior lobes. 2. This protein was detected in lysates of neurosecretory granules isolated from bovine pituitary posterior lobes. 3. The molecular weight appears to be close to 10000. 4. Neurophysin-C is similar in amino acid composition to neurophysin-I and -II; it contains a single residue of tyrosine and of methionine. The N-terminal amino acid in all three neurophysins is alanine. 5. Neurophysin-C accounts for approximately 15% of the total hormone-binding protein present in the pituitary posterior lobes. 6. The new neurophysin forms complexes with oxytocin as well as with [8-arginine]-vasopressin. The complex with vasopressin has been crystallized. 7. Bioassay of the pressor and oxytocic activities of the protein-hormone complexes shows that neurophysin-C binds one molecule of either vasopressin or oxytocin.

Acetone↗
PubMed1968

The isolation of the native hormone-binding proteins from bovine pituitary posterior lobes. Crystallization of neurophysin-I and-II as complexes with [8-arginine]-vasopressin.

1. The native hormone-binding proteins, neurophysin-I and -II, have been isolated from acetone-desiccated bovine pituitary posterior lobes. 2. Neurophysin-I and -II are present in approximately equal quantities in the tissue and are localized in the neurosecretory granules. 3. The apparent molecular weight, determined by equilibrium sedimentation of neurophysin-I, was 19000 and that of neurophysin-II was 21000; their sedimentation coefficients, S(20,w), were 1.66 and 2.02s respectively. 4. Neurophysin-I and -II are similar in amino acid composition. Neurophysin-II was distinguished from neurophysin-I by the absence of histidine. 5. The proteins form complexes with oxytocin as well as with vasopressin. Complexes of both proteins with [8-arginine]-vasopressin have been crystallized. 6. Bioassay of the pressor and oxytocic activities of the crystals shows that neurophysin-I binds three molecules of either vasopressin or oxytocin whereas neurophysin-II binds only two molecules of each hormone per molecule of protein. Complexes containing two molecules of oxytocin and one molecule of [8-arginine]-vasopressin per molecule of protein are formed by neurophysin-I and -II; both proteins appear to possess three polypeptide-binding sites/molecule.

Amino Acids↗
PubMed1967

Fractionation of neurophysin by molecular-sieve and ion-exchange chromatography.

Neurophysin has been separated into seven distinct protein fractions. One of these components had no hormone-binding activity. The fractions that had hormone-binding activity were similar in amino acid composition: their cystine content was in the range 11.5-14.5%. The major component, neurophysin-M, was distinguished from the protein isolated by van Dyke by the presence of methionine and the absence of histidine. Neurophysin-M binds both oxytocin and vasopressin with similar affinities.

Amino Acids↗
PubMed1967

The composition of crystalline complexes of neurophysin-M with [8-arginine]-vasopressin and oxytocin.

Neurophysin-M, a methionine-containing protein that is the major constituent of neurophysin, has been crystallized as complexes with [8-arginine]-vasopressin. Three moles of vasopressin alone or 2 moles of vasopressin together with 1 mole of oxytocin are bound/mole of protein. An amorphous complex of the protein with oxytocin alone contains 2 moles of the hormone/mole of protein. Deamino-[8-arginine]-vasopressin, a highly active basic analogue of vasopressin, is not bound by neurophysin. The primary amino group of both vasopressin and oxytocin is necessary for binding with neurophysin.

Journal Article↗
PubMedDate not supplied

Tyrosine kinase-mediated signal transduction pathways and the actions of polypeptide growth factors and G-protein-coupled agonists in smooth muscle.

This synopsis focuses on the role that tyrosine kinase pathways may play in the acute regulation of smooth muscle contractility by receptor-kinase-activating growth factors, such as epidermal growth factor-urogastrone (EGF-URO) and by G-protein-coupled agonists, such as angiotensin-II. Growth factor-activated response paradigms that modulate smooth muscle contractility are summarized and the parallels between the actions of G-protein-coupled agonists and growth factors in these response systems are pointed out. A possible dynamic interplay between tyrosine kinase and tyrosine phosphatase activities to modulate tissue tension is also hypothesized. Finally, a model is proposed, wherein an intermediary tyrosine kinase pathway is suggested as a point of convergence for the regulation of smooth muscle contractility by agonists as diverse as EGF-URO and angiotensin-II.

Angiotensin II↗
PubMedDate not supplied

Simultaneous phosphorylation of three human calpactins by kinase C.

We evaluated the concurrent phosphorylation of reconstituted mixtures of three purified human placental calpactins (or lipocortins) by purified bovine brain protein kinase C (PKC). Calpactin-I (p36 or lipocortin-II), calpactin-II (p38 or lipocortin-I), and a 70-kilodalton calpactin-related protein, calpactin-p70, when present together as substrates for PKC, all demonstrated comparable kinetic parameters (Vmax values = 0.3-0.5 nmol phosphate incorporated/min), with calpactin-II and calpactin-p70 exhibiting lower apparent Km values (40 and 30 nM, respectively) than did calpactin-I (Km, 200 nM). Because of the higher Vmax/Km ratios for calpactin-II and calpactin-70 (12.5 and 10.0, respectively) compared with the ratio for calpactin-I (2.0), our data suggest that, intracellularly, where all three calpactins might be co-localized, the higher molecular mass calpactins could be preferred substrates for PKC. Nonetheless, the requirement for relatively high calcium concentrations (greater than or equal to 0.5 mM) suggests that PKC-mediated phosphorylation of the calpactins may take place only in restricted intracellular compartments, wherein calcium concentrations might transiently reach levels much higher than those that are normally found intracellularly (less than or equal to 0.25 mM).

Annexins↗
PubMedDate not supplied

Radiolabelling of bovine myristoylated alanine-rich protein kinase C substrate (MARCKS) in an ADP-ribosylation reaction.

In an ADP-ribosylation reaction, we have observed the radiolabelling of a protein in a crude bovine brain homogenate, which upon two-dimensional gel electrophoresis migrated with an acidic pI (< 4.5) and an apparent molecular mass (80-90 kDa) consistent with the properties of the myristoylated, alanine-rich, protein kinase C substrate protein termed MARCKS. To establish the identity of this radiolabelled constituent in brain homogenates, we first purified bovine brain MARCKS using calmodulin-Sepharose affinity chromatography and we then supplemented the crude ADP-ribosylation reaction mixture with this purified MARCKS fraction. Concordant increases in radiolabelling and silver staining of the same protein component from the MARCKS-supplemented ADP-ribosylation reaction, as compared with the ADP-ribosylated crude homogenate, established the identity of this constituent as MARCKS. The radiolabelling of MARCKS was lower in comparison with the ADP-ribosylation of the related neuronal protein B-50/GAP-43 under identical reaction conditions. The potential functional consequences of the ADP-ribosylation of MARCKS are discussed and the possibility is raised that other members of the MARCKS family, such as the F52/MacMARCKS/MRP protein, may also be subject to ADP-ribosylation.

Adenosine Diphosphate Ribose↗
PubMedDate not supplied

Expression of platelet-derived growth factor receptor in rat heart allografts.

Growth factors may play a role in the tissue remodeling of long-surviving allografts. The aim of these experiments was to investigate the pattern of expression of platelet-derived growth factor and epidermal growth factor receptors in relation to the immunohistopathology of rat cardiac allografts. Heart grafts were transplanted between LEW and F344 and LEW.1A and LEW.1AR2 strains and removed for analysis at graft asystole or at predetermined time intervals. The grafts were studied with direct and indirect immunofluorescence and immunoperoxidase staining techniques and monoclonal antibodies against T lymphocytes, monocytes/macrophages, and endothelium. Expression of growth factor receptor was studied with the use of a monoclonal antibody against the platelet-derived growth factor-beta receptor and autoradiography with platelet-derived growth factor-BB or epidermal growth factor labeled with iodine 125. Increased expression of platelet-derived growth factor-beta receptors was observed in the media of graft arteries and in a number of myocardial cells in association with infiltration of the graft with T cells and monocytes/macrophages. Autoradiography with epidermal growth factor labeled with 125I failed to show changes in the binding pattern in relation to rejection. Upregulation of platelet-derived growth factor receptors may suggest involvement of platelet-derived growth factor in the tissue remodeling that occurs in conjunction with chronic rejection.

Animals↗