Institute for Clinical Chemistry and Laboratory Medicine

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Masjkur J.,TU Dresden | Arps-Forker C.,TU Dresden | Poser S.W.,TU Dresden | Nikolakopoulou P.,TU Dresden | And 12 more authors.
Journal of Biological Chemistry | Year: 2014

Background: The transcription factor Hes3 regulates the growth of neural and brain cancer stem cells.Results: Hes3 regulates growth, gene expression, evoked insulin release in cultured insulinoma cells, and sensitivity to streptozotocin in vivo.Conclusion: Hes3 is a novel regulator of cellular functions of importance in diabetes. ©2014 by The American Society for Biochemistry and Molecular Biology, Inc.


News Article | February 17, 2017
Site: www.futurity.org

Scientists discovered that a hemoglobin mutation was causing mild anemia in a young woman in Germany. But why did her father, who has the same mutation, not have anemia, too? The woman, who was in her 20s when diagnosed, and her father share a mutation in the gene that encodes hemoglobin, the protein in red blood cells responsible for taking up and delivering oxygen to cells around the body. The mutation is one of more than 1,000 discovered so far in adult human hemoglobin. Most appear to have no effect on people, but when medical problems occur, the disease is called a hemoglobinopathy and often named after the city or hospital where it was discovered. In this case, the family was living in Mannheim, Germany, but the father was born in the Turkish city of Kirklareli. The Kirklareli mutation did not affect the iron content of the father’s blood, but did appear to be the root cause of the young woman’s chronic anemia, researchers say. Further investigation revealed that he was a smoker—and his mutant hemoglobin was stabilized by carbon monoxide from the cigarettes he smoked. A paper on the research appears in the Journal of Biological Chemistry. The mutation is in the alpha subunit of human hemoglobin (H58L) and causes it to rapidly auto-oxidize, or rust, which causes the protein to fall apart, lose heme, and precipitate. As a result, the protein loses its ability to carry oxygen. Eventually, the red cells themselves become deformed and are destroyed, says John Olson, a biochemist at Rice University. Remarkably, this same mutation gives the protein an 80,000-fold higher affinity for carbon monoxide than for oxygen. Carbon monoxide from a cigarette will be selectively taken up by the mutant hemoglobin and prevent it from oxidizing and denaturing. This high affinity for carbon monoxide explained why the father showed no signs of anemia, Olson says. “He may never be an athlete because his blood can’t carry as much oxygen, but smoking has prevented him from being anemic. And there’s a side benefit. People with this trait are more resistant to carbon monoxide poisoning.” Olson says he does not know how or if the doctors treated the young woman. He doesn’t even know her name. But he suspected her iron-deficient anemia was more an annoyance than a threat to her life and would not recommend she start smoking to relieve it. “She shouldn’t smoke,” he says. “But she could take antioxidants, such as a lot of vitamin C, which would help prevent oxidation of her mutant hemoglobin. Her anemia is not that severe. At the same time, she shouldn’t worry too much about secondhand smoke, which might have a positive effect.” After ruling out common causes like blood loss, gastritis, or congenital defects, her doctors were curious enough about her ailment to call upon Emmanuel Bissé, a researcher at the Institute for Clinical Chemistry and Laboratory Medicine at the University of Freiburg, who discovered the mutation after sequencing her DNA. Bissé in turn recruited Olson and his team to help determine why the histidine-to-leucine change caused anemia in the daughter but not the father. Ironically, Ivan Birukou, a graduate student in Olson’s lab, had already generated the same mutation in human hemoglobin (one of several hundred made at Rice) to study how the protein rapidly and selectively binds oxygen. “Emmanuel wrote to me and said, ‘I know you’ve been making all these mutants in hemoglobin, and you’ve probably done the H58L mutation in (alpha) chains. Does this phenotype make sense?'” Olson recalls. “I said, ‘We can do a really neat study here, because we’ve already made the mutant hemoglobin in a recombinant system.’ We actually had a crystal structure (matching Kirklareli) that Ivan and (staff scientist) Jayashree Soman never published but had deposited in the Protein Data Bank. We had made this mutation to try to understand what the distal histidine was doing in alpha subunits.” They found in their 2010 study that replacing the histidine, which forms a strong hydrogen bond to oxygen, with leucine caused a dramatic decrease in oxygen affinity and an increase in carbon monoxide binding. Olson and Birukou realized back then that histidine played a key role in discriminating between oxygen and carbon monoxide in hemoglobin. “When Emmanuel wrote to me about his discovery, I already ‘knew’ what was happening with respect to carbon monoxide binding,” Olson says. The normal hydrogen bond causes bound oxygen to stick more tightly to hemoglobin in the same way hydrogen bonds cause spilled soda to feel sticky. “When you touch it, the sugar oxygens and hydrogens make hydrogen bonds with the polysaccharides on your finger,” Olson said. “That stickiness helps hold onto oxygen. But leucine is more like an oil, like butane or hexane, and oxygen does not stick well inside hemoglobin. In contrast, bound carbon monoxide is more like methane or ethane and can’t form hydrogen bonds.” Andres Benitez Cardenas, a postdoctoral researcher in Olson’s laboratory, did the crucial experiment in which he put carbon monoxide on the mutant alpha subunit of hemoglobin Kirklareli. The bound carbon monoxide slowed down oxidation of the protein and prevented loss of heme and precipitation. “In effect, Andres did the ‘smoking experiment’ to show why the father’s hemoglobin didn’t denature and cause anemia,” Olson said. The effect caused by Kirklareli, though unusual, is not unique. “There is another ‘smoking is good for you’ mutation,” he says, noting discoveries in Zurich in the late 1970s and early ’80s. That case mirrored the current collaboration, as the researchers looking for answers then sought help from Nobel Laureate Max Perutz, whose pioneering work on hemoglobin structures won him the prize in 1968. Olson himself served as a reviewer on some of the papers for hemoglobin Zurich in the 1980s. “Emmanuel knew that we had worked on these histidine-to-leucine mutations in myoglobin and hemoglobin, which is why he contacted us,” he says. “This type of collaboration is how science and medicine should work together.” Additional researchers from Rice, the University of Freiburg, and the University of Strasbourg are coauthors of the work.


Rice biochemist John Olson and collaborators in Germany and France helped a young woman and her father understand why she has anemia but her father, who is a smoker, does not. The woman, who was in her 20s when diagnosed, and her father share a mutation in the gene that encodes hemoglobin, the protein in red blood cells responsible for taking up and delivering oxygen to cells around the body. The mutation is one of more than 1,000 discovered so far in adult human hemoglobin. Most appear to have no effect on people, but when medical problems occur, the disease is called a hemoglobinopathy and often named after the city or hospital where it was discovered. In this case, the family was living in Mannheim, Germany, but the father was born in the Turkish city of Kirklareli. The Kirklareli mutation did not affect the iron content of her dad's blood, but did appear to be the root cause of the young woman's chronic anemia, according to the researchers. Further investigation revealed that absorbing carbon monoxide from cigarette smoke is therapeutic for those with this rare genetic disorder. A paper on the research appeared this month in the Journal of Biological Chemistry. The mutation is in the alpha subunit of human hemoglobin (H58L) and causes it to rapidly auto-oxidize, or rust, which causes the protein to fall apart, lose heme and precipitate. As a result, the protein loses its ability to carry oxygen. Eventually, Olson said, the red cells themselves become deformed and are destroyed. Remarkably, this same mutation gives the protein an 80,000-fold higher affinity for carbon monoxide than for oxygen. Carbon monoxide from a cigarette will be selectively taken up by the mutant hemoglobin and prevent it from oxidizing and denaturing. This high affinity for carbon monoxide explained why the father showed no signs of anemia, Olson said. "He may never be an athlete because his blood can't carry as much oxygen, but smoking has prevented him from being anemic," he said. "And there's a side benefit. People with this trait are more resistant to carbon monoxide poisoning." Olson said he does not know how or if the doctors treated the young woman. He doesn't even know her name. But he suspected her iron-deficient anemia was more an annoyance than a threat to her life and would not recommend she start smoking to relieve it. "She shouldn't smoke," he said. "But she could take antioxidants, such as a lot of vitamin C, which would help prevent oxidation of her mutant hemoglobin. Her anemia is not that severe. At the same time, she shouldn't worry too much about secondhand smoke, which might have a positive effect." After ruling out common causes like blood loss, gastritis or congenital defects, her doctors were curious enough about her ailment to call upon Emmanuel Bissé, a researcher at the Institute for Clinical Chemistry and Laboratory Medicine at the University of Freiburg, who discovered the mutation after sequencing her DNA. Bissé in turn recruited Olson and his team to help determine why the histidine-to-leucine change caused anemia in the daughter but not the father. Ironically, Ivan Birukou, a graduate student in Olson's lab, had already generated the same mutation in human hemoglobin (one of several hundred made at Rice) to study how the protein rapidly and selectively binds oxygen. "Emmanuel wrote to me and said, 'I know you've been making all these mutants in hemoglobin, and you've probably done the H58L mutation in (alpha) chains. Does this phenotype make sense?'" Olson recalled. "I said, 'We can do a really neat study here, because we've already made the mutant hemoglobin in a recombinant system.' We actually had a crystal structure (matching Kirklareli) that Ivan and (staff scientist) Jayashree Soman never published but had deposited in the Protein Data Bank. We had made this mutation to try to understand what the distal histidine was doing in alpha subunits." They found in their 2010 study that replacing the histidine, which forms a strong hydrogen bond to oxygen, with leucine caused a dramatic decrease in oxygen affinity and an increase in carbon monoxide binding. Olson and Birukou realized back then that histidine played a key role in discriminating between oxygen and carbon monoxide in hemoglobin. "When Emmanuel wrote to me about his discovery, I already 'knew' what was happening with respect to carbon monoxide binding," Olson said. He said that the normal hydrogen bond causes bound oxygen to stick more tightly to hemoglobin in the same way hydrogen bonds cause spilled soda to feel sticky. "When you touch it, the sugar oxygens and hydrogens make hydrogen bonds with the polysaccharides on your finger," Olson said. "That stickiness helps hold onto oxygen. But leucine is more like an oil, like butane or hexane, and oxygen does not stick well inside hemoglobin. In contrast, bound carbon monoxide is more like methane or ethane and can't form hydrogen bonds." Andres Benitez Cardenas, a postdoctoral researcher in Olson's laboratory, did the crucial experiment in which he put carbon monoxide on the mutant alpha subunit of hemoglobin Kirklareli. The bound carbon monoxide slowed down oxidation of the protein and prevented loss of heme and precipitation. "In effect, Andres did the 'smoking experiment' to show why the father's hemoglobin didn't denature and cause anemia," Olson said. He said the effect caused by Kirklareli, though unusual, is not unique. "There is another 'smoking is good for you' mutation," he said, noting discoveries in Zurich in the late 1970s and early '80s. That case mirrored the current collaboration, as the researchers looking for answers then sought help from Nobel Laureate Max Perutz, whose pioneering work on hemoglobin structures won him the prize in 1968. Olson himself served as a reviewer on some of the papers for hemoglobin Zurich in the 1980s. "Emmanuel knew that we had worked on these histidine-to-leucine mutations in myoglobin and hemoglobin, which is why he contacted us," he said. "This type of collaboration is how science and medicine should work together." Explore further: Protect against carbon monoxide as researchers hunt antidote More information: Emmanuel Bissé et al. Hemoglobin Kirklareli (α H58L), a New Variant Associated with Iron Deficiency and Increased CO Binding, Journal of Biological Chemistry (2017). DOI: 10.1074/jbc.M116.764274


News Article | February 16, 2017
Site: www.eurekalert.org

There's at least one person in the world for whom smoking has a beneficial effect, and it took an international collaboration of scientists led by a Rice University professor to figure out why. Rice biochemist John Olson and collaborators in Germany and France helped a young woman and her father understand why she has anemia but her father, who is a smoker, does not. The woman, who was in her 20s when diagnosed, and her father share a mutation in the gene that encodes hemoglobin, the protein in red blood cells responsible for taking up and delivering oxygen to cells around the body. The mutation is one of more than 1,000 discovered so far in adult human hemoglobin. Most appear to have no effect on people, but when medical problems occur, the disease is called a hemoglobinopathy and often named after the city or hospital where it was discovered. In this case, the family was living in Mannheim, Germany, but the father was born in the Turkish city of Kirklareli. The Kirklareli mutation did not affect the iron content of her dad's blood, but did appear to be the root cause of the young woman's chronic anemia, according to the researchers. Further investigation revealed that absorbing carbon monoxide from cigarette smoke is therapeutic for those with this rare genetic disorder. A paper on the research appeared this month in the Journal of Biological Chemistry. The mutation is in the alpha subunit of human hemoglobin (H58L) and causes it to rapidly auto-oxidize, or rust, which causes the protein to fall apart, lose heme and precipitate. As a result, the protein loses its ability to carry oxygen. Eventually, Olson said, the red cells themselves become deformed and are destroyed. Remarkably, this same mutation gives the protein an 80,000-fold higher affinity for carbon monoxide than for oxygen. Carbon monoxide from a cigarette will be selectively taken up by the mutant hemoglobin and prevent it from oxidizing and denaturing. This high affinity for carbon monoxide explained why the father showed no signs of anemia, Olson said. "He may never be an athlete because his blood can't carry as much oxygen, but smoking has prevented him from being anemic," he said. "And there's a side benefit. People with this trait are more resistant to carbon monoxide poisoning." Olson said he does not know how or if the doctors treated the young woman. He doesn't even know her name. But he suspected her iron-deficient anemia was more an annoyance than a threat to her life and would not recommend she start smoking to relieve it. "She shouldn't smoke," he said. "But she could take antioxidants, such as a lot of vitamin C, which would help prevent oxidation of her mutant hemoglobin. Her anemia is not that severe. At the same time, she shouldn't worry too much about secondhand smoke, which might have a positive effect." After ruling out common causes like blood loss, gastritis or congenital defects, her doctors were curious enough about her ailment to call upon Emmanuel Bissé, a researcher at the Institute for Clinical Chemistry and Laboratory Medicine at the University of Freiburg, who discovered the mutation after sequencing her DNA. Bissé in turn recruited Olson and his team to help determine why the histidine-to-leucine change caused anemia in the daughter but not the father. Ironically, Ivan Birukou, a graduate student in Olson's lab, had already generated the same mutation in human hemoglobin (one of several hundred made at Rice) to study how the protein rapidly and selectively binds oxygen. "Emmanuel wrote to me and said, 'I know you've been making all these mutants in hemoglobin, and you've probably done the H58L mutation in (alpha) chains. Does this phenotype make sense?'" Olson recalled. "I said, 'We can do a really neat study here, because we've already made the mutant hemoglobin in a recombinant system.' We actually had a crystal structure (matching Kirklareli) that Ivan and (staff scientist) Jayashree Soman never published but had deposited in the Protein Data Bank. We had made this mutation to try to understand what the distal histidine was doing in alpha subunits." They found in their 2010 study that replacing the histidine, which forms a strong hydrogen bond to oxygen, with leucine caused a dramatic decrease in oxygen affinity and an increase in carbon monoxide binding. Olson and Birukou realized back then that histidine played a key role in discriminating between oxygen and carbon monoxide in hemoglobin. "When Emmanuel wrote to me about his discovery, I already 'knew' what was happening with respect to carbon monoxide binding," Olson said. He said that the normal hydrogen bond causes bound oxygen to stick more tightly to hemoglobin in the same way hydrogen bonds cause spilled soda to feel sticky. "When you touch it, the sugar oxygens and hydrogens make hydrogen bonds with the polysaccharides on your finger," Olson said. "That stickiness helps hold onto oxygen. But leucine is more like an oil, like butane or hexane, and oxygen does not stick well inside hemoglobin. In contrast, bound carbon monoxide is more like methane or ethane and can't form hydrogen bonds." Andres Benitez Cardenas, a postdoctoral researcher in Olson's laboratory, did the crucial experiment in which he put carbon monoxide on the mutant alpha subunit of hemoglobin Kirklareli. The bound carbon monoxide slowed down oxidation of the protein and prevented loss of heme and precipitation. "In effect, Andres did the 'smoking experiment' to show why the father's hemoglobin didn't denature and cause anemia," Olson said. He said the effect caused by Kirklareli, though unusual, is not unique. "There is another 'smoking is good for you' mutation," he said, noting discoveries in Zurich in the late 1970s and early '80s. That case mirrored the current collaboration, as the researchers looking for answers then sought help from Nobel Laureate Max Perutz, whose pioneering work on hemoglobin structures won him the prize in 1968. Olson himself served as a reviewer on some of the papers for hemoglobin Zurich in the 1980s. "Emmanuel knew that we had worked on these histidine-to-leucine mutations in myoglobin and hemoglobin, which is why he contacted us," he said. "This type of collaboration is how science and medicine should work together." Bissé is lead author of the paper. Co-authors are Christine Schaeffer-Reiss, Alain Van Dorsselaer and Tchilabalo Dilezitoko Alayi of the University of Strasbourg, France, and the Hubert CURIEN Multidisciplinary Institute, Strasbourg; Thomas Epting and Karl Winkler of the Institute for Clinical Chemistry and Laboratory Medicine at the University of Freiburg; and Birukou, Benitez Cardenas, Soman and graduate student Premila Samuel at Rice. Birukou is now a technical expert at Syngenta Crop Protection, North Carolina. Olson is the Ralph and Dorothy Looney Professor of Biochemistry and Cell Biology at Rice. This news release can be found online at http://news. Located on a 300-acre forested campus in Houston, Rice University is consistently ranked among the nation's top 20 universities by U.S. News & World Report. Rice has highly respected schools of Architecture, Business, Continuing Studies, Engineering, Humanities, Music, Natural Sciences and Social Sciences and is home to the Baker Institute for Public Policy. With 3,879 undergraduates and 2,861 graduate students, Rice's undergraduate student-to-faculty ratio is 6-to-1. Its residential college system builds close-knit communities and lifelong friendships, just one reason why Rice is ranked No. 1 for happiest students and for lots of race/class interaction by the Princeton Review. Rice is also rated as a best value among private universities by Kiplinger's Personal Finance. To read "What they're saying about Rice," go to http://tinyurl. .


PubMed | University of Greifswald, Interfaculty Institute for Genetics and Functional Genomics, University of Duisburg - Essen, University of Lübeck and Institute for Clinical Chemistry and Laboratory Medicine
Type: Journal Article | Journal: European thyroid journal | Year: 2015

Hyperthyroidism is known to induce a hypercoagulable state. It stimulates plasma levels of procoagulative factors and reduces fibrinolytic activity. So far most of the data have been derived from patients with endogenous hyperthyroidism with a wide variability in the underlying pathogenesis and severity of the disease.In this study we experimentally induced thyrotoxicosis in healthy volunteers to explore the effects of thyroxine excess on the plasma proteome. Using a shotgun proteomics approach, the abundance of plasma proteins was monitored before, during and after thyrotoxicosis.Sixteen healthy male subjects were sampled at baseline, 4 and 8 weeks under 250 g/day thyroxine p.o., as well as 4 and 8 weeks after stopping the application. Plasma proteins were analyzed after depletion of 6 high-abundance proteins (MARS6) by LC-ESI-MS/MS mass spectrometry. Mass spectrometric raw data were processed using a label-free, intensity-based workflow. Subsequently, the linear dependence between protein abundances and fT4 levels were calculated using a Pearson correlation.All subjects developed biochemical thyrotoxicosis, and this effect was reversed within the first 4 weeks of follow-up. None of the volunteers noticed any subjective symptoms. Levels of 10 proteins involved in the coagulation cascade specifically correlated with fT4, supporting an influence of thyroid hormone levels on blood coagulation even at nonpathological levels.The results suggest that experimental thyrotoxicosis exerts selective and specific thyroxine-induced effects on coagulation markers. Our study design allows assessment of thyroid hormone effects on plasma protein levels without secondary effects of other diseases or therapies.


Wanninger J.,University of Regensburg | Liebisch G.,Institute for Clinical Chemistry and Laboratory Medicine | Schmitz G.,Institute for Clinical Chemistry and Laboratory Medicine | Bauer S.,University of Regensburg | And 5 more authors.
Experimental and Molecular Pathology | Year: 2012

Adiponectin protects from hepatic fat storage but adiponectin deficient mice (APN-/-) fed a standard chow do not develop liver steatosis. This indicates that other pathways might be activated to compensate for adiponectin deficiency. An unbiased and comprehensive screen was performed to identify hepatic alterations of lipid classes in these mice. APN-/- mice had decreased hepatic cholesteryl esters while active SREBP2 and systemic total cholesterol were not altered. Upregulation of cytochromes for bile acid synthesis suggests enhanced biliary cholesterol excretion. Analysis of 37 individual fatty acid species showed reduced stearate whereas total fatty acids were not altered. Total amount of triglycerides and phospholipids were equally abundant. A selective increase of monounsaturated phosphatidylcholine and phosphatidylethanolamine which positively correlate with hepatic and systemic triglycerides with the latter being elevated in APN-/- mice, was identified. Stearoyl-CoA desaturase 1 (SCD1) is involved in the synthesis of monounsaturated fatty acids and despite higher mRNA expression enzyme activity was not enhanced. Glucosylceramide postulated to contribute to liver damage was decreased. This study demonstrates that adiponectin deficiency is associated with hepatic changes in lipid classes in mice fed a standard chow which may protect from liver steatosis. © 2012 Elsevier Inc. All rights reserved.


PubMed | Institute for Clinical Chemistry and Laboratory Medicine
Type: Journal Article | Journal: Transfusion | Year: 2014

Stored platelet (PLT) concentrates (PLCs) for transfusion develop a PLT storage lesion (PSL), decreasing PLT viability and function with profound lipidomic changes and PLT extracellular vesicle (PL-EV) release. High-density lipoprotein 3 (HDL3 ) improves PLT homeostasis through silencing effects on PLT activation in vivo. This prompted us to investigate HDL3 and apolipoprotein A-I (apoA-I) as PSL-antagonizing agents.Healthy donor PLCs were split into low-volume standard PLC storage bags and incubated with native (n)HDL3 or apoA-I from plasma ethanol fractionation (precipitate IV) for 5 days under standard blood banking conditions. Flow cytometry, Born aggregometry, and lipid mass spectrometry were carried out to analyze PL-EV release, PLT aggregation, agonist-induced PLT surface marker expression, and PLT and plasma lipid compositions.Compared to control, added nHDL3 and apoA-I significantly reduced PL-EV release by up to -62% during 5 days, correlating with the added apoA-I concentration. At the lipid level, nHDL3 and apoA-I antagonized PLT lipid loss (+12%) and decreased cholesteryl ester (CE)/free cholesterol (FC) ratios (-69%), whereas in plasma polyunsaturated/saturated CE ratios increased (+3%) and CE 16:0/20:4 ratios decreased (-5%). Administration of nHDL3 increased PLT bis(monoacylglycero)phosphate/phosphatidylglycerol (+102%) and phosphatidic acid/lysophosphatidic acid (+255%) ratios and improved thrombin receptor-activating peptide 6-induced PLT aggregation (+5%).nHDL3 and apoA-I improve PLT membrane homeostasis and intracellular lipid processing and increase CE efflux, antagonizing PSL-related reduction in PLT viability and function and PL-EV release. We suggest uptake and catabolism of nHDL3 into the PLT open canalicular system. As supplement in PLCs, nHDL3 or apoA-I from Fraction IV of plasma ethanol fractionation have the potential to improve PLC quality to prolong storage.

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