scholarly journals Structural determinants and genetic modifications enhance BMP2 stability and extracellular secretion

2018 ◽  
Author(s):  
Vinayak Khattar ◽  
Joo Hyoung Lee ◽  
Hong Wang ◽  
Soniya Bastola ◽  
Selvarangan Ponnazhagan
Keyword(s):  
Half Life ◽  
Proteasomal Degradation ◽  
Structural Determinants ◽  
Extracellular Secretion ◽  
Short Half Life ◽  
Genetic Modifications ◽  
Lysine Residues ◽  
Stability Results ◽  
Enhanced Stability

ABSTRACTThe short half-life and use of recombinant bone morphogentic protein (BMP)-2 in large doses poses major limitations in the clinic. Events regulating post-translational processing and degradation of BMP2 in situ, linked to its secretion, have not been understood. Towards identifying mechanisms regulating intracellular BMP2 stability, we first discovered that inhibiting proteasomal degradation enhances both intracellular BMP2 level and its extracellular secretion. Next, we identified BMP2 degradation occurs through an ubiquitin-mediated mechanism. Since ubiquitination precedes proteasomal turnover and mainly occurs on lysine residues of nascent proteins, we systematically mutated individual lysine residues within BMP2 and tested them for enhanced stability. Results revealed that substitutions on four lysine residues within the pro-BMP2 region and three in the mature region increased both BMP2 turnover and extracellular secretion. Structural modeling revealed key lysine residues involved in proteasomal degradation occupy a lysine cluster near proprotein convertase cleavage site. Interestingly, mutations within these residues did not affect biological activity of BMP2. These data suggest preventing intracellular proteasomal loss of BMP2 through genetic modifications can overcome limitations related to its short half-life.

scholarly journals Structural determinants and genetic modifications enhance BMP2 stability and extracellular secretion

2019 ◽  
Vol 1 (3) ◽  
pp. 180-190
Author(s):  
Vinayak Khattar ◽  
Joo Hyoung Lee ◽  
Hong Wang ◽  
Soniya Bastola ◽  
Selvarangan Ponnazhagan
Keyword(s):  
Structural Determinants ◽  
Extracellular Secretion ◽  
Genetic Modifications

Kinetics of Various 99mTc-Sn-Pyrophosphate Compounds in the Rat

1977 ◽  
Vol 16 (04) ◽  
pp. 157-162 ◽  
Author(s):  
C. Schümichen ◽  
B. Mackenbrock ◽  
G. Hoffmann
Keyword(s):  
Normal Saline ◽  
Half Life ◽  
Compound A ◽  
Short Half Life ◽  
Low Concentrations ◽  
The Stability ◽  
Kinetics Of ◽  
In Vitro Stability

SummaryThe bone-seeking 99mTc-Sn-pyrophosphate compound (compound A) was diluted both in vitro and in vivo and proved to be unstable both in vitro and in vivo. However, stability was much better in vivo than in vitro and thus the in vitro stability of compound A after dilution in various mediums could be followed up by a consecutive evaluation of the in vivo distribution in the rat. After dilution in neutral normal saline compound A is metastable and after a short half-life it is transformed into the other 99mTc-Sn-pyrophosphate compound A is metastable and after a short half-life in bone but in the kidneys. After dilution in normal saline of low pH and in buffering solutions the stability of compound A is increased. In human plasma compound A is relatively stable but not in plasma water. When compound B is formed in a buffering solution, uptake in the kidneys and excretion in urine is lowered and blood concentration increased.It is assumed that the association of protons to compound A will increase its stability at low concentrations while that to compound B will lead to a strong protein bond in plasma. It is concluded that compound A will not be stable in vivo because of a lack of stability in the extravascular space, and that the protein bond in plasma will be a measure of its in vivo stability.

scholarly journals Tafazzins from Drosophila and mammalian cells assemble in large protein complexes with a short half-life

Mitochondrion ◽  
2015 ◽  
Vol 21 ◽  
pp. 27-32 ◽  
Author(s):  
Yang Xu ◽  
Ashim Malhotra ◽  
Steven M. Claypool ◽  
Mindong Ren ◽  
Michael Schlame
Keyword(s):  
Half Life ◽  
Mammalian Cells ◽  
Protein Complexes ◽  
Large Protein ◽  
Short Half Life

scholarly journals In-Situ Cosmogenic 14C: Production and Examples of its Unique Applications in Studies of Terrestrial and Extraterrestrial Processes

Radiocarbon ◽  
2001 ◽  
Vol 43 (2B) ◽  
pp. 731-742 ◽  
Author(s):  
D Lal ◽  
A J T Jull
Keyword(s):  
Half Life ◽  
Earth Science ◽  
Chemical Properties ◽  
Radioactive Isotopes ◽  
Earth’S Atmosphere ◽  
The Earth ◽  
Wide Range ◽  
History Of ◽  
Earth's Atmosphere

Nuclear interactions of cosmic rays produce a number of stable and radioactive isotopes on the earth (Lai and Peters 1967). Two of these, 14C and 10Be, find applications as tracers in a wide variety of earth science problems by virtue of their special combination of attributes: 1) their source functions, 2) their half-lives, and 3) their chemical properties. The radioisotope, 14C (half-life = 5730 yr) produced in the earth's atmosphere was the first to be discovered (Anderson et al. 1947; Libby 1952). The next longer-lived isotope, also produced in the earth's atmosphere, 10Be (half-life = 1.5 myr) was discovered independently by two groups within a decade (Arnold 1956; Goel et al. 1957; Lal 1991a). Both the isotopes are produced efficiently in the earth's atmosphere, and also in solids on the earth's surface. Independently and jointly they serve as useful tracers for characterizing the evolutionary history of a wide range of materials and artifacts. Here, we specifically focus on the production of 14C in terrestrial solids, designated as in-situ-produced 14C (to differentiate it from atmospheric 14C, initially produced in the atmosphere). We also illustrate the application to several earth science problems. This is a relatively new area of investigations, using 14C as a tracer, which was made possible by the development of accelerator mass spectrometry (AMS). The availability of the in-situ 14C variety has enormously enhanced the overall scope of 14C as a tracer (singly or together with in-situ-produced 10Be), which eminently qualifies it as a unique tracer for studying earth sciences.

Pulmonary vasodilation by nitric oxide gas and prodrug aerosols in acute pulmonary hypertension

1998 ◽  
Vol 84 (2) ◽  
pp. 435-441 ◽  
Author(s):  
Christophe Adrie ◽  
Fumito Ichinose ◽  
Alexandra Holzmann ◽  
Larry Keefer ◽  
William E. Hurford ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Pulmonary Hypertension ◽  
Vascular Resistance ◽  
Pulmonary Circulation ◽  
Half Life ◽  
Hemodynamic Effects ◽  
Pulmonary Vasodilation ◽  
Short Half Life ◽  
Acute Pulmonary Hypertension ◽  
Inhaled No

Adrie, Christophe, Fumito Ichinose, Alexandra Holzmann, Larry Keefer, William E. Hurford, and Warren M. Zapol. Pulmonary vasodilation by nitric oxide gas and prodrug aerosols in acute pulmonary hypertension. J. Appl. Physiol. 84(2): 435–441, 1998.—Sodium 1-( N, N-diethylamino)diazen-1-ium-1,2-diolate {DEA/NO; Et2N[N(O)NO]Na} is a compound that spontaneously generates nitric oxide (NO). Because of its short half-life (2.1 min), we hypothesized that inhaling DEA/NO aerosol would selectively dilate the pulmonary circulation without decreasing systemic arterial pressure. We compared the pulmonary selectivity of this new NO donor with two other reference drugs: inhaled NO and inhaled sodium nitroprusside (SNP). In seven awake sheep with pulmonary hypertension induced by the infusion of U-46619, we compared the hemodynamic effects of DEA/NO with those of incremental doses of inhaled NO gas. In seven additional awake sheep, we examined the hemodynamic effects of incremental doses of inhaled nitroprusside (i.e., SNP). Inhaled NO gas selectively dilated the pulmonary vasculature. Inhaled DEA/NO produced nonselective vasodilation; both systemic vascular resistance (SVR) and pulmonary vascular resistance (PVR) were reduced. Inhaled SNP selectively dilated the pulmonary circulation at low concentrations (≤10−2 M), inducing a decrease of PVR of up to 42% without any significant decrease of SVR (−5%), but nonselectively dilated the systemic circulation at larger doses (>10−2 M). In conclusion, despite its short half-life, DEA/NO is not a selective pulmonary vasodilator compared with inhaled NO. Inhaled SNP appears to be selective to the pulmonary circulation at low doses but not at higher levels.

Identification of absorption lines of short half-life actinides in the spectrum of Przybylski’s star (HD 101065)

2008 ◽  
Vol 24 (2) ◽  
pp. 89-98 ◽  
Author(s):  
V. F. Gopka ◽  
A. V. Yushchenko ◽  
V. A. Yushchenko ◽  
I. V. Panov ◽  
Ch. Kim
Keyword(s):  
Half Life ◽  
Absorption Lines ◽  
Short Half Life

Synthetic Strategies for the Modification of Diclofenac

Synlett ◽  
2017 ◽  
Vol 28 (15) ◽  
pp. 1984-1989 ◽  
Author(s):  
Rudolf Schneider ◽  
Stephan Schmidt ◽  
Sven Hanelt ◽  
Carsten Canitz ◽  
Holger Hoffmann ◽  
...  
Keyword(s):  
Amino Acid ◽  
Solid Phase ◽  
Free Acid ◽  
Protein A ◽  
Sensor Applications ◽  
Protein Conjugates ◽  
Lysine Residues ◽  
Sufficient Distance ◽  
Direct Binding

For many heterogeneous sensor applications as well as the synthesis of hapten antigens to produce antibodies, protein conjugates of the target substance are essential. A requirement is that the target substance already offers or is modified to contain a functionality that allows for coupling to a protein, that is, an amino acid residue. Ideally, to avoid shielding of the compound by the carrier protein, a sufficient distance to the protein surface should be provided. With its carboxyl function diclofenac (DCF) allows for direct binding to lysine residues after in situ synthesis of the NHS ester. One problem is that diclofenac as free acid tends to autocondensation, which results in low yields. Here we describe the ‘insertion’ of a C6 spacer via synthesis of the amide with 6-aminohexanoic acid. To carry out the reaction in solution, first the methyl ester of the amino acid had to be produced. Due to otherwise low yields and large cleaning efforts, solid-phase synthesis on Fmoc Ahx Wang resin is recommended. The crude product is mainly contaminated by cleavage products from the resin which were removed by chromatography. The structure of the highly pure hapten was completely determined by nuclear magnetic resonance (NMR) spectroscopy.

scholarly journals A phosphoinositide-binding cluster in cavin1 acts as a molecular sensor for cavin1 degradation

2015 ◽  
Vol 26 (20) ◽  
pp. 3561-3569 ◽  
Author(s):  
Vikas A. Tillu ◽  
Oleksiy Kovtun ◽  
Kerrie-Ann McMahon ◽  
Brett M. Collins ◽  
Robert G. Parton
Keyword(s):  
Proteasomal Degradation ◽  
Resting Cells ◽  
Coat Proteins ◽  
Cellular Processes ◽  
Molecular Sensor ◽  
Lysine Residues ◽  
Low Levels ◽  
Membrane Stretching ◽  
Signaling Modules ◽  
Site Release

Caveolae are abundant surface organelles implicated in a range of cellular processes. Two classes of proteins work together to generate caveolae: integral membrane proteins termed caveolins and cytoplasmic coat proteins called cavins. Caveolae respond to membrane stress by releasing cavins into the cytosol. A crucial aspect of this model is tight regulation of cytosolic pools of cavin under resting conditions. We now show that a recently identified region of cavin1 that can bind phosphoinositide (PI) lipids is also a major site of ubiquitylation. Ubiquitylation of lysines within this site leads to rapid proteasomal degradation. In cells that lack caveolins and caveolae, cavin1 is cytosolic and rapidly degraded as compared with cells in which cavin1 is associated with caveolae. Membrane stretching causes caveolar disassembly, release of cavin complexes into the cytosol, and increased proteasomal degradation of wild-type cavin1 but not mutant cavin1 lacking the major ubiquitylation site. Release of cavin1 from caveolae thus leads to exposure of key lysine residues in the PI-binding region, acting as a trigger for cavin1 ubiquitylation and down-regulation. This mutually exclusive PI-binding/ubiquitylation mechanism may help maintain low levels of cytosolic cavin1 in resting cells, a prerequisite for cavins acting as signaling modules following release from caveolae.

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