
Most people know about single nucleotide polymorphisms (SNPs) or single amino acid polymorphisms (SAPs), but few people ever get told how they operate in health and disease.
N-acyl amino acids in humans can involve acetylation, and C-terminus amino acids can be involved with methylation. Both are controlled by light. This post describes how the process happens.
Modifying the C-terminus of proteins is essential in understanding what light is doing biophysically to determine what biochemistry is possible in a cell. The C-terminus is also known as the carboxyl-terminus, carboxy-terminus, C-terminal tail, C-terminal end, or COOH-terminus. It represents the end of an amino acid chain (protein or polypeptide), terminated by a free carboxyl group (-COOH). When the protein is translated from messenger RNA, it is created from the N-terminus to the C-terminus. The convention for writing peptide sequences is to put the C-terminal end on the right and write the sequence from N- to C-terminus.
For example, while the N-terminus of a protein often contains targeting signals, the C-terminus can include retention signals for protein sorting. The most common Endoplasmic Reticulum retention signal is the amino acid sequence -KDEL (Lys-Asp-Glu-Leu) or -HDEL (His-Asp-Glu-Leu) at the C-terminus. This keeps the protein in the endoplasmic reticulum and prevents it from entering the secretory pathway in a cell.
For example, when the skin over the gut with a massive amount of POMC is not stimulated by solar light, POMC is lightly translated, and Vitamin D levels stay low. As a result, one form of C-terminal modification is prenylation. During prenylation, a farnesyl- or geranylgeranyl-isoprenoid membrane anchor is added to a cysteine residue near the C-terminus. Small molecular weight, membrane-bound G proteins are often modified this way. The circadian mechanism in cells operates via G proteins. With proper solar light, prenylation goes smoothly.
The enterocytes in the gut form a barrier. They include a mechanical boundary against pathogens, antigens, and toxins; the monolayer of intestinal epithelial cells (IECs) represents the human body’s largest contact area with the environment due to villi folding. From their early development in the crypt bottom until the shedding of aged cells at the villus tip, IECs follow a continuous turnover process. This process is controlled by the circadian mechanism in the SCN and transmitted to the gut via the peripheral nervous system and the melanin sheets in the organ of Zuckerlandl, where melanin and POMC reside.
Under physiological conditions, a complex interaction between cytoskeleton rearrangement and tight junction proteins guarantees that the cell shedding itself does not mean a disturbance of epithelial integrity. However, alterations of epithelial integrity lead to the development of gut inflammatory disorders, such as inflammatory bowel diseases (IBDs). IBDs are associated with marked alterations of IECs, leading to increased tight junction permeability, altered cytoskeletal rearrangement, and induction of epithelial cell death with a subsequent loss of barrier function. All IBDs are linked to poor solar exposure and or artificial light at night on the skin or via the eyes.
In Crohn’s disease, there is Rho-A prenylation, and the absent solar light then changes protein signaling in the enterocytes of the gut. This links epithelial homeostasis directly to intestinal inflammation.
The peptide created by POMC before cleavage is composed of three such segments: N-POMC, which is located at the N-terminus; ACTH, which is located in the middle; and β-LPH, which is located at the C-terminus. Each of these segments contains one MSH sequence: γ-MSH in N-POMC, α-MSH in ACTH, and β-MSH in β-LPH.
The N-terminus is also known as the amino-terminus, NH2-terminus, N-terminal end, or amine-terminus, is the start of a protein or polypeptide, referring to the free amine group (-NH2) located at the end of a polypeptide. Look at the structure of melanin below when it interacts with an electrophile metal.
When a protein is translated from messenger RNA, it is created from the N-terminus to the C-terminus. The amino end of an amino acid (on a charged tRNA), during the elongation stage of translation, attaches to the carboxyl end of the growing chain. Since the start codon of the genetic code codes for ONLY one amino acid, methionine, most protein sequences start with a methionine (or, in bacteria, mitochondria, and chloroplasts, the modified version N-formylmethionine, fMet). I wrote a blog on that topic on Patreon. You should have expected I told you this for a reason before the melanin story came out of my mitochondrial mist. The circadian mechanism operates 100% of the time in a posttranslational fashion. I laid out that case at the Palestra Health conference when I presented to several politicians and hedge fund managers who were present. Some proteins are modified posttranslationally, for example, by cleavage from a protein precursor and therefore may have different amino acids at their N-terminus. Light frequencies can change the N-terminus in proteins.
The N-terminus is the first part of the protein that exits the ribosome during protein biosynthesis. It often contains signal peptide sequences, “intracellular postal codes,” that direct the delivery of the protein to the proper organelle. The signal peptide is typically removed at the destination by a signal peptidase. The N-terminal amino acid of a protein is an essential determinant of its half-life. This is how the ubiquitin system operates, and quantum mechanics raises or lessens the probability of the likelihood of a protein being degraded. This is called the N-end rule.
The N-terminal signal peptide is recognized by the signal recognition particle (SRP) and results in the targeting of the protein to the secretory pathway. In eukaryotic cells, these proteins are synthesized at the rough endoplasmic reticulum. Protein N-termini can be modified co – or post-translationally. Modifications include the removal of initiator methionine (iMet) by aminopeptidases, attachment of small chemical groups such as acetyl, propionyl, and methyl, and the addition of membrane anchors, such as palmitoyl and myristoyl groups. There are many other N-terminus modifications like methylation. Light frequencies dictate how biochemistry unfolds.
N-terminus modification is how many diseases begin with aberrant light choices. Many pharmacological, genetic, and mechanistic studies in mammals have suggested that certain members of the N-acyl amino acids stimulate mitochondrial respiration and whole-body energy expenditure directly. Other complementary studies have also established roles for N-acyl amino acids in glucose homeostasis, adipogenesis, vascular tone, and bone homeostasis. This explains thoroughly why POMC has the protein construction it does.
The N-POMC is located at the N-terminus ACTH, which is away from the N and C terminus because it is located in the middle of the protein. This is a deep evolutionary clue about how mammals used ACTH to create sugar from light cleavage. What should shock you is that each of these cleavage segments contains one MSH sequence: γ-MSH in N-POMC, α-MSH in ACTH, and β-MSH in β-LPH. This tells you that light and dark are controlling the peptide’s cleavage.
Notably, the functional consequence and enzymatic regulation of each N-acyl amino acid in mammals highly depend on the structural properties of the fatty acid tail group and amino acid head group.
By integrating whole genome sequencing data with N-acyl amino acid levels, one can identify the EPIGENETIC determinants of N-acyl amino acid levels and cluster according to the amino acid head group. For example, in mammalian heart disease, this is why CYP4F2 is associated with many human cardiometabolic disorders. Centralized science has now identified the CYP4F2 locus as an epigenetic determinant of plasma N-oleoyl-leucine and N-oleoyl-phenylalanine levels in human plasma, as the papers below show. In experimental studies, it has now been demonstrated that CYP4F2-mediated hydroxylation of N-oleoyl-leucine and N-oleoyl-phenylalanine results in metabolic diversification and production of many previously unknown lipid metabolites in humans with varying characteristics of the fatty acid tail group, including several that structurally resemble fatty acid hydroxy fatty acids. These studies provide a structural framework for understanding the regulation and disease associations of N-acyl amino acids in humans and identify that the diversity of this lipid signaling family can be significantly expanded through CYP4F-mediated ω-hydroxylation.
Modern “centralized chemical biology” relies increasingly on protein chemistry, which ideally allows precise positioning of labels, cargoes, and post-translational modifications (PTMs) in the contexts of complex protein structures. The resulting modified proteins prove helpful in determining proper physiology and Big Pharma therapeutic applications. They allow us to understand the decentralized cell by the probing and modulating function, as well as their tracking and (un)caging in cells that happen with light and dark cycles.
Big Pharma chemists have developed various methods to control the convergent construction of site-selectively modified proteins. Traditionally, the non-site-selective chemical modification of proteins has relied on the nucleophilicity of the side-chains of natural amino acid residues like lysine (Lys) and cysteine (Cys), as well as protein N-terminus through direct acylation, alkylation and arylation with a wide array of electrophiles. In chemistry, an electrophile is a chemical species that forms bonds with nucleophiles by accepting an electron pair. Electrophiles accept electrons in biochemistry. Biological electrophiles are defined as electron-deficient species of chemicals that include heavy metals, environmental pollutants, toxic drug metabolites, flavor enhancers like Splenda, cell signaling mediators, and unsaturated aldehyde products of membrane lipid peroxidation.
When DHA is in the SN-2 position, it becomes planar. This is what turns the central retinal pathways of the retinohypothalamic tracts into a wide band gapped semiconductive LED array that targets melanin all over the brain. Melanin absorbs all light, ROS, and RNS to charge separate water to create a tremendous amount of electrons to power the system. In this illustration, electrons act as a canvas does in a painting: sunlight is the paint, and electrons are the brushes.
SUMMARY
BLUE LIGHT IS A STIMULANT THAT CREATES ROS/RNS NORMALLY
And stimulants are 👍 great. Most Americans drink a few cups of stimulants each morning ☕️ to get themselves up to face the daily grind.
But you know what’s not great? Being stimulated 24 hours of every day by blue light. This ruins the dose-response curve of the ROS/RNS. That is exactly what is occurring to modern humans because they live indoors and use tech screens excessively. What else is different about this version of blue light? The blue light that wakes us up in the sun is NEVER present without 42% IR-A light, which is red. AM sunlight has 42% red light and only 1600K of blue light. This small stimulus of blue light is about to improve executive function of the prefrontal cortex. This blue light needs red light to control the oxidation ROS/RNS that blue light makes when present without red light in our light environment.
ALL CELLS contain ion channels in their outer (plasma) and inner (organelle) membranes. Ion channels, similar to other proteins, are targets of oxidative impact, which modulates ion fluxes across membranes. Subsequently, these ion currents affect electrical excitability, such as action potential discharge (in neurons, muscle, and receptor cells), alteration of the membrane resting potential, synaptic transmission, hormone secretion, muscle contraction, or coordination of the cell cycle. Circadian biology controls all of this.
An important class of ion channels is the family of potassium (K+) channels (in the nitric oxide cycle); they are not only in charge of the membrane resting potential or the repolarization of the action potentials but also control cell proliferation or transmitter/hormone release, to name a few. A subgroup of K+ channels are the so-called calcium (Ca2+) activated K+ channels, which need either an increase of Ca2+ at their intracellular face to open or a combination of Ca2+ and voltage to function correctly. Maxi Ca2+-activated K+ channels, also named BK channels, constitute a subgroup of Ca2+-activated K+ channels.
Do you know where these ion channels exist in humans? They are found on the inner mitochondrial membrane. EXCESSIVE BLUE LIGHT exposure destroys these potassium ion channels to ruin signaling of cells that control the circadian mechanism and are associated with leptin and melanopsin. LET THAT SINK IN with all the ways modification happens on the C and N terminus of proteins. I only shared a couple with you in this blog.
Mitochondria are a significant source of ROS generation targeting BK channels. Blue light creates that stimulus when RED LIGHT IS ABSENT.
The inner membrane of mitochondria contains BK channels (mtBK), which appear essential in the production of ROS. mtBK channels appear to be inserted into the mitochondrial membrane with the toxin binding sites for charybdotoxin and iberiotoxin exposed to the mitochondrial intermembrane space. This can be accessed using outside-out patch configuration of the inner mitochondrial membrane. Consequently, the C-terminal tail domain, including the Ca2+ binding site, is localized to the mitochondrial matrix where the proton gradient exists.
RED LIGHT MOVES PROTONS BEST. Blue light creates the most ROS. Do you understand why subtracting red and UV light from blue creates mitochondrial diseases now?
Can you imagine what the centralized chemists controlled by Big Pharma may find when they look at C and N terminus modifications by light frequencies in POMC or DHA in humans?
CITES
1. Turner JR. Intestinal mucosal barrier function in health and disease. Nat Rev Immunol. 2009;9(11):799–809.View this article via: PubMed CrossRef Google Scholar
2. van der Flier LG, Clevers H. Stem cells, self-renewal, and differentiation in the intestinal epithelium. Annu Rev Physiol. 2009;71:241–260. View this article via: PubMed CrossRef Google Scholar
3. Watson AJ, Duckworth CA, Guan Y, Montrose MH. Mechanisms of epithelial cell shedding in the Mammalian intestine and maintenance of barrier function. Ann N Y Acad Sci. 2009;1165:135–142.
4. Long J.Z., Svensson K.J, Bateman L.A., Lin H., Kamenecka T., Lokurkar I.A. et al. The secreted enzyme PM20D1 regulates lipidated amino acid uncouplers of mitochondria. Cell. 2016; 166: 424-435
5. https://www.cell.com/cell-chemical-biology/abstract/S2451-9456(20)30146-X