The fast and the furious phosphor. Its action in innate immunity

The fast and the furious phosphor: Its action in innate immunity.

Friedrich Flachsbart

H. Minasyan discovered the antimicrobial action of erythrocytes.¹

F. Flachsbart discussed with him the antimicrobial action of coagulation and the action of unfractionated heparin in infection, especially sepsis.²

But the question remains:

Is there something like a deeper common ground of innate immunity?

Prof. Goebel showed in microbes that the metabolism is connected with invasive behaviour and that mammalian metabolism is connected with defence of microbial invasion.³

Is metabolism also the common ground of different lines of defence against invasion?

Let us try to find an answer by looking back to the beginning of life on earth.

When life on earth began, it was a combination of phosphor, sugar and nitrogen.

These elements build up the RNA world. This term was first used 1986 by Gilbert.⁴

Phosphor, the sugar ribose and a nucleobase combine to RNA.

RNA is the most important ingredient of life, especially in the “constant battle between infectious agents trying to subvert host defence systems and host cells protecting themselves from infection”.⁵ It started with RNA. And it still runs with RNA.

The question is:

How are these first ingredients of life still integrated in the communication between different species of microbial and eukaryotic species?

A first answer is:

They are still the first line of interaction between microbe and man.

Let us start with phosphor.

Phosphor provides energy. Phosphorylation activates other molecules.

Phosphor acts as information transmitter. Phosphorylation opens channels that provide information to other molecules, even cascades and nets of molecules. The enzymes of phosphorylation are called kinases. Phosphatases stop their action. Kinases and phosphatases form a highly integrated network of information flow. Edwin H. Fischer and Edwin G. Krebs showed us this fascinating world of phosphorylation.⁶

Phosphor is also the backbone of ribonucleic acid, RNA, the older nucleic acid.

RNA is very active in transformation and translation of the genetic DNA-information.

RNA is also part of many regulations and actions of cells.

There are important differences between the four RNA-molecules. But all four are part of the innate immunity, at different places and structures. Let us find the differences between the four ribonucleotides.

All four ribonucleotides attach phosphor and bind it in the four RNA-Triphosphates:

Adenosine-Tri-Phosphate (ATP).

Guanosine-Tri-Phosphate (GTP).

Uridine-Tri-Phosphate (UTP).

Cytidine-Tri-Phosphate (CTP).

1. ATP and GTP store energy.

ATP, Adenosine-Tri-Phosphate, is the most important energy transporter.

ATP and its derivatives NADPH, FADH store energy and transmit energy.

GTP, Guanosine-Tri-Phosphate is also an energy transporter.

GTP receives energy mostly from the citric-acid-cycle.

2. ATP and GTP transmit information.

ATP also serves as signal molecule.

The binding of phosphate by enzymes, called kinases, is the starter of enzyme action. Phosphor is a regulatory molecule, Phosphorylation induces the action of many enzymes.

Phosphatases stop this action. Kinases and Phosphatases are combined in a fine tuned network.

Phosphorylation by kinases means „go“, dephosphorylation by phosphatases means „stop“.

3. GTP acts with GPCR: Transmembrane information flow.

GTP is the substrate of G-PCR, G-Protein-Coupled-Receptors. Seven-helical-transmembrane-receptors are inactive with GTP. Dephosphorylation of GTP to GDP changes the protein-conformation and the information from outside the membrane is transformed into the action of number of signal-molecules inside the cell. These GPCR are involved in many information-pathways. Most pharmacological substances act via GPCR. But the most important function may be the interaction with microbes.⁷

4. Signalling with cyclic Monophosphates. Second messengers.

Adenyl cyclase forms cyclo-AMP (c-AMP), Guanyl cyclase build cyclo-GMP (c-GMP). These are important signal-molecules.⁸ They are triggered by small molecules, hormones, neurotransmitters like adrenalin, serotonin. Together with Calcium 2+, 1,2-Diacylglycerol (DG) and Inositol-1,4,5-Triphosphat (IP3) they activate the Protein-Kinase and Protein-Phosphatase-Networks. In fungi cAMP induces pathogenic structures.⁹ In Mycobacterium tuberculosis cAMP is upregulated in the state of infection.¹⁰ cAMP is active in the pathogen and host, cAMP can be a helper or a killer, friend or foe.¹¹ cAMP protects bacteria against viral infections,¹² cAMP protects man against virus.¹³ It was found, that heparin interacts with cAMP.¹⁴

5. UTP is the coenzyme for activation of sugar.

The phosphorylation of sugar allows the building of polymers like glycogen. The Glucose-1-Phosphate-UTP-Transferase changes UTP and glucose-1-phosphat into an UDP-hexose and pyrophosphate. UDP-glucose builds glycogen, the sugar-store. Connections between UTP, glycogen and microbes begin to emerge.¹⁵

6. CTP is part of membrane-building.

CTP and phosphocholine transforms into CDP-choline and pyrophosphate. CDP-diacylglycerol (DG) and phosphatidylinositol are the signal molecules together with Ca2+, cAMP and cGMP, as written above. Phosphatidylcholine is the outer membrane molecule, 50 % of the membrane are phosphatidylcholine. Base-exchange from phosphatidyl-choline and serine build up choline and phosphatidylserine, 10 % of the phospholipid of man. Phosphatidylserine is most important in the inside-out flippase of membranes.¹⁶ Phosphatidylserine seems to be a central focus of sepsis induced organ dysfunction.¹⁷

7. Phosphate and haemoglobin. (2,3-DPG)

A stereochemical change of haemoglobin is triggered by a small molecule, 2,3-Diphosphoglycerat (2,3-DPG) changes the oxygen-transport-capacity of haemoglobin. Without 2,3-DPG haemoglobin has only 8 % oxygen-transport-capacity. 2,3-DPG could also be a defence against reactive oxygen species (ROS) and reactive nitrogen species (NOS).¹⁸

8. Phosphate, Glucose and haemoglobin. (Favism)

The stereochemical change of glucose is induced by phosphor. Phosphorylation of glucose to glucose-6-phosphat is the starting point of glycolysis.¹⁹ Glucose is not able to leave the cell. In erythrocytes energy is provided only by glucose. The mutation of the enzyme glucose-6-phosphate-dehydrogenase²⁰ induces a haemolytic anaemia.²¹ This disease, called favism since Hippocrates, is a survival benefit against malaria (like thalassaemia and sickle-cell-anaemia). Sickle cell anaemia is a major cause of thrombosis.²²

9. Phosphate and Diabetes.

2002 the idea was presented: Silent occult streptococci are the common soil of diabetes and vascular disease.²³ The kinase-cascades of IKKbeta induce an inflammation in rheumatic fever and in insulin-resistance. Aspirin (7g/day) stops the activation of the kinases.²⁴ The effect of aspirin was 1877 first documented by Wilhelm Ebstein, who later became director of the internal medicine clinic of the university Goettingen. 1901 his observation was again documented by Williamson.²⁵ 1957 rheumatic fever and diabetes were together treated with one drug, aspirin.²⁶

10. Phosphate and Gout/Pseudogout.

The end product of ribonucleic-acid (nitrogenous base-pentose-phosphate) is uric acid, inducing gout.²⁷ Pseudogout is the association of phosphate with calcium.²⁸ Gout and pseudogout are inflammatory states. They are antimicrobial inflammations, uric acid and calcium-phosphate kill the microbes. Prof. Wilhelm Ebstein, Göttingen, wondered more than 100 years ago, why there was not one microbe found in gout, when there were millions of streptococci visible in erysipelas in his patients.²⁹

11. Phosphate, Xanthin and Asthma.

Adenosin and ATP interact with allergens to induce asthma. The receptors are part of a purinergic signalling system.³⁰ Heparin is a therapeutic antagonist of severe chemical induced bronchial destruction.³¹

12. PolyP, the polyanion unfractionated heparin and RNA, DNA, histones and NET act in thrombosis.

Polyphosphates (PolyP) are stored in dense granules within thrombocytes. PolyP triggers coagulation Factor XII (Hagemann-Factor).³² Bradykinin is part of an inflammatory/thrombotic pathway.³³

Polyphosphates (PolyP) and unfractionated heparin are stored in granules within mast cells.³⁴ It is suggested, „that the release of heparin coupled with polyP inhibits the procoagulant properties of polyP while retaining the proinflammatory capability.³⁵

Polyphosphates and unfractionated heparin are polyanions.³⁶ They do not fit into the concept of „key and lock“: They activate other molecules not only by a specific key that does fit into a specific hole.³⁷ PolyP and UFH activate other molecules by diffuse interactions of many atoms of the molecules.

Extracellular RNA is another polyanion that interferes with coagulation like PolyP.³⁸ Both induce thrombosis. Unfractionated Heparin is the antagonist, inducing bleeding.

DNA, histones and NET, neutrophil extracellular traps, also interact with coagulation. All three are in complex ways intertwined with coagulation, induce thrombosis.³⁹ Unfractionated heparin is the antagonist, inducing bleeding.

UFH binds to heparin-binding-proteins (HBP) in an unspecific manner. And there are hundreds of proven UFH-Protein-combinations.⁴⁰ LMWH, interacting with a specific pentasaccharide of a thrombosis-factor seems to be not inferior in prevention of venous thrombosis. But UFH does so much more, it interacts with membranes, proteins, cells. In the USA only UFH is allowed in heart-surgery and dialysis.

UFH interacts with thrombosis. UFH interacts earlier in the road to infection, in the attachment of a pathogen. Ebola is an example. UFH stops the entrance of the virus. LMWH, interacting with thrombosis, does not interact with the virus-entry.⁴¹

This is the answer to the question: “Why is the sepsis mortality so high?”⁴²

Sepsis mortality became so high, because UFH was lost in the treatment of sepsis. Only in extreme thrombotic states UFH is allowed.⁴³

Unfractionated heparin interacts with invading pathogens in various protein-combinations. UFH dissolves attachment of pathogens to eukaryotic cells. Prof. Kuhn started his therapy of septic abortion in 1964 with 30.000 IU/day UFH one hour before antibiotic treatment. And so he could prevent intravascular thrombosis of kidney and lung, disseminated intravascular coagulation and bleeding.⁴⁴ UFH does have many different contact-phases, not one as LMWH and not two, as Levi wrote.⁴⁵ Sepsis and thrombocytopenia can be changed by the polyanion UFH, not by LMWH.⁴⁶

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