Category Archives: Aargh ! Big pharma sheds chemists. Why?

Action at a distance comes to chemistry

Allostery is an abstract concept in protein chemistry, far removed from everyday life. Far removed except if you like to breathe, or have ever used a benzodiazepine (Valium, Librium, Halcion, Ativan, Klonopin, Xanax) for anything. Breathing? Really? Yes — Hemoglobin, the red in red blood cells is really 4 separate proteins bound to each other. Each of the four can bind one oxygen molecule. Binding of oxygen to one of the 4 proteins produces a subtle change in the structure of the other 3, making it easier for another oxygen to bind. This produces another subtle change in structure of the other making it easier for a third oxygen to bind. Etc.

This is what allostery is, binding of molecule to one part of a protein causing changes in structure all over the protein.

Neurologists are familiar with the benzodiazepines, using them to stop continuous seizure activity (status epilepticus), treat anxiety (Xanax), or seizures (Klonopin). They all work the same way, binding to a complex of 5 proteins called the GABA receptor, which when it binds Gamma Amino Butyric Acid (GABA) in one place causes negative ions to flow into the neuron, inhibiting it from firing. The benzodiazepines bind to a completely different site, making the receptor more likely to open when it binds GABA.

The assumption about all allostery is that something binds in one place, pushing the atoms around, which push on other atoms which push on other atoms, until the desired effect is produced. This is the opposite of action at a distance, where an effect is produced without the necessity of physical contact.

Even though Newton invented a theory of gravity, which worked beautifully, he was disturbed by the fact that it acted through empty space. Here’s what he wrote in a letter to Bentley

“That gravity should be innate inherent & {essential} to matter so that one body may act upon another at a distance through a vacuum without the mediation of any thing else by & through which their action or force {may} be conveyed from one to another is to me so great an absurdity that I beleive no man who has in philosophical matters any competent faculty of thinking can ever fall into it. “

So physicists invented the ether which was physical, and allowed objects to push each other around by pushing on the ether between them.

But action at a distance without one atom pushing on the next etc. etc. is exactly what an incredible paper found [ Proc. Natl. Acad. Sci. vol. 117 pp. 25445 – 25454 ’20 ]. Here’s a link but it’s probably behind a paywall —

The paper studied TetR, a protein containing 203 amino acids. If you’ve ever thought about it, almost all the antibiotics we have come from bacteria, which they use on other bacteria. Since we still have bacteria around, the survivors must have developed a way to resist antibiotics, and they’ve been doing this long before we appeared on the scene.

TetR helps bacteria resist tetracycline, an antibiotic produced by bacteria. When tetracycline binds to TetR it causes other parts of the protein to change so it binds DNA causing the bacterium, among other things, to make a pump which moves tetracyline out of the cell. Notice that site where tetracycline binds on TetR is not the business end where TetR binds DNA, just as where the benzodiazepines bind the GABA receptor is not where the ion channel is.

This post is long enough already without describing the cleverness which allowed the authors to do the following. They were able to make TetRs containing every possible mutation of all 203 positions. How many is that — 203 x 19 = 3838 different proteins. Why 19? Because we have 20 amino acids, so there are 19 possible distinct changes at each of the 203 positions in TetR.

Some of the mutants didn’t bind to DNA, implying they were non-functional. The 3 dimensional structure of TetR is known, and they chose 5 of nonfunctional mutants. Interestingly these were distributed all over the protein.

Then, for each of the 5 mutants they made another 3838 mutants, to see if a mutation in another position would make the mutant functional again. You can see what a tremendous amount of work this was.

Here is where it gets really interesting. The restoring mutant (revertants if you want to get fancy) were all over the protein and up to 40 – 50 Angstroms away from the site of the dead mutation. Recall that 1 Angstrom is the size of a hydrogen atom, a turn of the alpha helix is 5.4 Angstroms and contains 3.5 amino acids per turn.The revertant mutants weren’t close to the part of the protein binding tetracycline or the part binding to DNA.

Even worse the authors couldn’t find a contiguous path of atom pushing atom pushing atom, to explain why TetR was able to bind DNA again. So there you have it — allosteric action at a distance.

There is much more in the paper, but after all the work they did it’s time to let the authors speak for themselves. “Several important insights emerged from these results. First, TetR exhibits a high degree of allosteric plasticity evidenced by the ease of disrupting and restoring function through several mutational paths. This suggests the functional landscape of al- lostery is dense with fitness peaks, unlike binding or catalysis where fitness peaks are sparse. Second, allosterically coupled residues may not lie along the shortest path linking allosteric and active sites but can occur over long distances “

But there is still more to think about, particularly for drug development. Normally, in developing a drug for X, we have a particular site on a particular protein as a target, say the site on a neurotransmitter receptor where a neurotransmitter binds. But the work shows that sites far removed from the actual target might have the same effect

Why drug development is hard #34 — designer hallucinogens

NBOMe (2-(4-Bromo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl) methyl]ethanamine to you) is a potent hallucinogen, a member of the phenylethylamine series of hallucinogens.  Well that’s the same as saying the current Intel chips are a member of the Intel class of starting with the 8080. has the structure, but I count 2 methoxy groups and a bromine on the phenyl group and a methoxy benzyl group making the amine group a secondary amine.

How anyone came up with the structure will remain unknown to me as it was part of a PhD thesis written in 2003 — unfortunately in German —Ralf Heim (February 28, 2010). “Synthese und Pharmakologie potenter 5-HT2A-Rezeptoragonisten mit N-2-Methoxybenzyl-Partialstruktur. Entwicklung eines neuen Struktur-Wirkungskonzepts.” (in German). Retrieved 2013-05-10.

Like other hallucinogens (LSD, mescaline, psilocin) NBOMe binds to the 2A variety of serotonin receptor (aka 5HT2A — at least 16 serotonin receptors are known) and acts like LSD as an agonist.

Which brings me to Cell vol. 182 pp. 1574 – 1588 ’20 —, probably behind a paywall.  Which has beautiful cryoEM structures of 5HT2A bound to LSD, NBOMe and methiothepin, an inverse agonist.  To get pictures they had to stabilize the structure with a single chain variable fragment of an antibody (something that always makes me wonder how physiologic the structure obtained actually is).

Why use NBOMe as an example of how hard drug discovery is?  Well the binding site of LSD to 5HT2A is well known, and the paper has some beautiful pictures of LSD snuggled between the 7 transmembrane segments of 5HT2A.  What is remarkable about NBOMe is that it lies in the binding site in a completely different orientation.  Moreover NBOMe fits in a previously undescribed pocket between transmembrane segments #3 and #6 (TM3, TM6).  Actually I think NBOMe actually produces the pocket.

So even if you know the target of your drug (5HT2A) and how another drug hits the target you’re aiming for, this doesn’t help you in designing a newer and more potent drug.

The Gordian knot untied, the undruggable surface drugged

The claim is made that the surface of a coiled coil is flat hence undruggable because there are no nooks and crannies for a drug to fit in.  Well the surface of a coiled coil isn’t really flat but  PNAS vol. 117 pp. 17195 – 17203 ’20 did actually drug one such structure using FKBP12 to help it.

Coiled coil containing proteins are interesting.  There are roughly 3.5 amino acids per turn of each alpha helix, so to get a repeating structure 7 amino acids are required.  They are numbered a through g from amino to carboxy.  Positions a and d are usually hydrophobic amino acids (Leu, Ile, Val, Ala), e and g are usually polar and charged.  The non polar a and d side chains associate with the opposite alpha helix by knobs into holes packing.  Each individuual helix is right handed but the two helices wind around each other with a right handed turn.  Actually it’s a bit more complicated — there are 3.64 amino acids/turn (not 3.5), so to get the number of amino acids down so there are 3.5 per turn (allowing the structure to repeat every 7 amino acids, left handed supercoiling of each helix occurs.  No question, this is an elegant design.  Pauling may have figured out the alpha helix, but I doubt that anyone proposed the coiled coil before it was actually discovered — does anyone out there know?

All intermediate filament proteins (neurofilaments, vimentin, keratin) contain a coiled coil structure.

CEP250 is a large protein (2,442 amino acids) found in the microtubule organizing center (centrosome).  It has a lot of long alpha helices wound around each other.  The picture they show has two alpha helices (each with 24 or so turns) wound around each other — that’s what a coiled coil is.

The authors used a small (12 kiloDalton) widely distributed protein (FKBP12) to accomplish this.  FKBP12 is important because it can complex the immunosuppressants, rapamycin and FK506 allowing them to bind to their targets (mTOR and calcineurin respectively).  Basically FKBP12 forms a torus with the coiled coil inside, with the drug at the hole in the torus where it is held against the coiled coil — clever work.

This isn’t to deny the difficulty of drugging a flat surface which unfortunately characterize most protein protein interfaces.


How can a cytokine still act after it’s gone

Here is yet another new wrinkle in how drugs act.

Type I interferons are  very well studied cytokines, important in the defense against viruses.  There are 16 of them, and in response to infection, they induce the expression of over 300 antiviral genes (ISGs — Interferon Stimulated Genes).

Type I interferon is gone from the plasma after intramuscular injection in a few hours, yet its effects last days to weeks.  Well maybe the proteins it induces hang around a long time.

Not so says Proc. Natl. Acad. Sci. vol. 117 pp. 17510 – 17512 ’20.  It hangs around inside the cell in little vesicles formed from pinching off part of the cell membrane containing type I interferon bound to its receptor.  These are called endosomes.  The authors call this siloing.  The silos persist for days and can actually be seen if you use fluorescently labeled type I interferon.  The endosomes with type I interferon inside persist for days.

I’m sure people are actively studying other cytokines of which there are many, many — e.g. there are more than 50 interleukins that we know about.

Why drug development is hard #32 and #33

The bloodbath among drug chemists continues (see Derek’s recent posts — because drug development is very hard and success is rare. Two nearly back to back papers in PNAS show just how hard drug development is (and why).

Animal models of human disease have a poor track record in pointing to new drugs.  One reason is that humans have new genes that animals don’t. One example is the horribly named CHRFAM7A, a dominant negative inhibitor of the alpha7 nicotinic cholinergic receptor [ PNAS vol. 116 pp. 7932 – 7940 ’16 ].

Alpha7 is found on macrophages where it exerts an anti-inflammatory action. Alpha7 agonists work beautifully in rodent inflammatory disease models.  They crashed and burned in human trials.  Why?  Because CHRFAM7A  binds to Alpha7 blocking the ability of acetyl choline to bind to it.  It is a totally new gene for man. It arose when 5 exons of the UL kinase 4 gene on chromosome #3 translocated nd then fused with the Dupa gene, which itself originated with 5 exons partially duplicated from the 10 exon alpha7 gene on the forward strand of chromosome #15.  So CHRFAM7A in close proximity to alpha7 (about which much more in the next post) and structurally similar to it.

[ PNAS vol. 116 pp. 7957 – 7962 ’19 ] Practically next door is a paper about MI-2, a drug thought to be useful in a (fortunately) rare brain tumor of childhood — diffuse intrinsic pontine glioma (maybe 3 cases in 38 years of practice).  Menin is a tumor suppressor lacking in a less rare syndrome (Type I Multiple Endocrine Neoplasia). MI-2 inhibits menin, but this paper shows that this isn’t its mechanism of action. Rather it inhibits an enzyme on the biosynthetic route to cholesterol (lanosterol synthetase).  So even when you think you know what a drug should be doing (which is probably why MI-2 was developed), that may not be how it works.

Why drug development is hard #31: retroviruses at the synapse

What if I told you that a very important neuronal synaptic protein Arc (Arg3.1) is acting like like a virus, sending copies of itself (and its messenger RNA) across the synapse?  Would a team of shrinks, who’ve never examined me, tell you that I was crazy and unfit to blog?  Well there is very good evidence that exactly this occurs in one situation and probably many more [ Cell vol. 172 pp. 8 – 10, 262 – 274, 275 – 288 ’18] —

Arc stands for Activity Regulated Cytoskeleton associated protein.  It’s messenger RNA (mRNA) is transcribed from the gene in response to neuronal activity.  More importantly, the mRNA for  Arc is rapidly distributed to active synapses through the cell body and dendrites, where it is translated into protein. It is locally and rapidly stimulated during the induction of long term depression and plays a critical role in removing a class of glutamic acid receptors (AMPA receptors) from the synapse.  To whet the interest of drug developers, Arc regulates the activity dependent cleavage of the Amyloid Precursor Protein (APP) and beta amyloid production by its interaction with presenilin

Several posts could easily be filled with what Arc does, but that’s not what is so amazing about these papers.  Parts of the Arc protein arose from one of the many transcriptionally dead retroviruses found in our genome.  Our species literally wouldn’t exist without other retroviral gifts.  For instance syncytin1 is a protein expressed a high levels in the placenta.  It is produced from the envelope gene of an endogenous retrovirus (HERV-W) which has undergon inactivating mutations in its other major genes (gag and pol).  Mutant mice in which the gene has been knocked out die in utero due to failure of placenta formation.

Part of the arc gene arose from the Gag gene (Group specific antigen gene) of a retrovirus.  Recall most viruses have proteins coating their genetic material when they’re on the move (e. g. a capsid).  In the case of retroviruses, the genetic material is RNA rather than DNA.  Well the gag elements of the Arc protein form a capsid containing the mRNA for Arc (just like a virus).  In some way or other the capsid containing mRNA gets outside the neuron at the nerve muscle junction and gets into muscle.  The evidence is good that this happens, but in a system somewhat removed from us — the fruitfly (Drosophila).  Fruitfly neuromuscular junctions lacking this mechanism are weaker.

Well that’s pretty far from us.  However one of the papers (275 – 288) showed that the Arc protein and its mRNA was found in extracellular vesicles released from mouse neurons cultured from their cerebral cortex.  Could viral-like particles be crossing the synapses in our brains (which are already pretty chockfull of stuff — see  It’s very early times (in fact the Cell issue came out 3 days ago) but people are sure to look.  There are at least 100 Gag derived genes in the human genome (Campillos, M., Doerks, T., Shah, P.K., and Bork, P. (2006). Computational characterization of multiple Gag-like human proteins. Trends Genet. 22, 585–589.).

Remarkable.  Remember CRISPR was hiding in plain sight for half a century.  We have a lot to learn.  No wonder drugs have unexpected side effects.

Why drug development is hard #30 — more new interactions we had no idea existed

We’re full of proteins which bind RNA wrangling it into a desired conformation.  The ribosome (whose enzymatic business end is pure RNA) has a mere 80 proteins doing this.  Its mass is 4,300,000 times that of a hydrogen atom.  However the idea that RNA could return the favor was pretty much unheard of until [ Science vol. 358 pp. 993 – 994, 1051 – 1055 ’17 — ].

As is often the case, viruses and the RNA world continue to instruct us.  In order to survive, some viruses induce cells to express a long (2,200+ nucleotides) nonCoding (for protein that is) RNA called lncRNA-ACOD1.   It binds to a protein enzyme (called GOT2, for Glutamic acid OxaloAcetic Transaminase 2) increasing its catalytic efficiency.  This shifts cellular metabolism around making it more favorable for virus proliferation, as GOT2 is found in mitochondria being used to replenish tricarboxylic cycle intermediates — e.g. making more energy available to the virus.

lncRNA-ACOD1 is induced by a variety of viruses, most importantly influenza virus in man, and vaccinia, herpes simplex 1, vesicular stomatitis virus in mice.  Exactly how viruses induce it isn’t clear, but the transcription factor NFkappaB is involved.

Viruses continue to teach us.  The amino acids of GOT2 (#15 – #68) and the interacting sequence of nucleotides in lncRNA-ACOD1 (#165 – #390) are well conserved across species.  This might be a primordial mechanism from the RNA world (forgotten but not gone) to produce ATP production to compe with metabolic stress.   The RNA/protein binding site is close (4.2 Angstroms) to the substrate binding site.

The fun is just starting as several other lncRNAs are induced by viruses.  You can only imagine what they will tell us.  Another set of drug targets perhaps, or worse, the cause of peculiar side effects from drugs already in use.

Why drug discovery is hard #29 — a very old player doing a very new thing

We all know what RNA does don’t we?  It binds to other RNAs and to DNA.  Sure lots of new forms of RNA have been found: microRNAs, competitive endogenous RNA (ceRNA), long nonCoding (for protein) RNA (lncRNA), piwiRNAs, small interfering RNAs (siRNAs), . .. The list appears endless.  But the basic mechanism of action of RNA in the cell is binding to some other polynucleotide (RNA or DNA) and affecting its function.

Not so fast.  A new paper describes  lncRNA-ACOD1, a cellular RNA induced by a variety of viruses.  lncRNA-ACOD1 binds to an enzyme enhancing its catalytic efficiency.  Now that’s new.  Certainly RNAs and proteins bind to each other in the ribosome, and in RNAase P, but here the proteins serve to structure the RNA so it can carry out its catalytic function, not the other way around.

The enzyme bound is called GOT2 (Glutamic Oxaloacetic Transaminase 2).  Much interesting cellular biochemistry is discussed in the paper which I’ll skip, except to say that the virus uses the hyped up GOT2 to repurpose the cell’s metabolic machinery for its own evil ends.

lncRNA-ACOD1 has 3 exons and a polyAdenine tail.  There are two transcript variants containing  2,330 and 2,259 nucleotides.  There are only 100 copies/cell.  lncRNA-ACOD1 nucleotides #165 – #390 bind to amino acids #54 – #68 of GOT2.

So what are the other 2000 or so nucleotides of lncRNA-ACOD1 doing?   The phenomenon of RNA binding to protein is quite likely to be more widespread.  Both the GOT2 interacting motif and the interacting sequence of lncRNA-ACOD1 are well conserved across species of hosts and viruses.

Although viruses co-opt lncRNA-ACOD1, it is normally expressed in the heart as is GOT2 with no viral infection at all.  So we have likely stumbled onto an entirely new method of cellular metabolic control, AND a whole new set of players and interactions for drugs to act on (if they aren’t already doing this unknown to us).

This is series member #29 of why drug development is hard, most of which concentrated on the fact that we don’t know all the players.  lncRNA-ACOD1 is different — RNA is a player we’ve known for a very long time  but it appears to be playing a game entirely new to us.

It is also good to see cutting edge research like this coming out of China.  Hopefully it will stand up, but enough questionable stuff has come from them that every Chinese paper is under a cloud.

This is why I love reading the current literature.  You never know what you’re going to find.  It’s like opening presents.

A possible new player

Drug development is very hard because we don’t know all the players inside the cell. A recent paper describes an entirely new class of player — circular DNA derived from an ancient virus.  The authoress is Laura Manuelidis, who would have been a med school classmate had I chosen to go to Yale med instead of Penn.   She is the last scientist standing who doesn’t believe Prusiner’s prion hypothesis.  She didn’t marry the boss’s daughter being female, so she married the boss instead;  Elias Manuelidis a Yale neuropathologist who would be 99 today had he not passed away at 72 in 1992.

The circular DNAs go by the name of SPHINX  an acronym  for  Slow Progressive Hidden INfections of X origin.  They have no sequences in common with bacterial or eukaryotic DNA, but there some homology to a virus infecting Acinebacter, a wound pathogen common in soil and water.

How did she find them?  By doggedly pursuing the idea the neurodegenerative diseases such as Cruetzfeldt Jakob Disease (CJD) and scrapie were due to an infectious agent triggering aggregation of the prion protein.

As she says:  “The cytoplasm of CJD and scrapie-infected cells, but not control cells, also contains virus-like particle arrays and because we were able to isolate these nuclease-protected particles with quantitative recovery of infectivity, but with little or no detectable PrP (Prion Protein), we began to analyze protected nucleic acids. Using Φ29 rolling circle amplification, several circular DNA sequences of <5 kb (kilobases) with ORFs (Open Reading Frames) were thereby discovered in brain and cultured neuronal cell lines. These circular DNA sequences were named SPHINX elements for their initial association with slow progressive hidden infections of X origin."

SPHINX itself codes for a 324 amino acid protein, which is found in human brain, concentrated in synaptic boutons.  Strangely, even though the DNAs are presumably viral derived, they contain intervening sequences which don't code for protein.

The use of rolling circle amplification is quite clever, as it will copy only circular DNA.

Stanley Prusiner is sure to weigh in.  Remarkably, Prusiner was at Penn Med when I was and was even in my med school fraternity (Nu Sigma Nu)  primarily a place to eat lunch and dinner.  I probably ate with him, but have no recollection of him whatsoever.

Circular DNAs outside chromosomes are called plasmids. Bacteria are full of them. The best known eukaryote containing plasmids is yeast. Perhaps we have them as well. Manuelidis may be the first person to look.

What is docosahexenoic acid and why should you care?

Why should drug chemists care about docosahexenoic acid — it’s a fairly trivial organic structure as these things go – a 22 carbon straight chain carboxylic acid with 6 double bonds — However the structure is decidedly non-random (see later)

Docosahexenoic acid turns out to be crucial for the function of the blood brain barrier (BBB), something that makes it very difficult to get drugs into the brain. Years of work have shown that the only drugs able to get through the BBB are small lipid soluble molecules of mass under 400 kiloDaltons with fewer than 9 hydrogen bonds. Certainly not a large group of drugs. The more we know about the BBB, the more likely we’ll be able to figure out something to circumvent it.

The BBB was known to exist more than 100 years ago. Ehrlich found that dyes injected into the circulation were rapidly taken up by all organs except the brain. His student E. Goldmann found that dye injected into the CSF stained the brain but not other organs.

The barrier has at least two components — (1) a very tight seal between the cells lining brain blood vessels (e.g. the endothelium) — see the end of the post — (2)very low transfer across the endothelial cell from the vessel lumen. The latter is called transcytosis and involves formation of small vesicles at the lumenal surface of the endothelium, migration across the endothelial cell with release of vesicle content on the other side.

In general there are two mechanisms of transcytosis — clathrin coated pits, and caveolae. Brain endothelium shows very low rates of transcytosis. There aren’t any coated pits (no explanation I can find) and the rate of caveolar transcytosis is very low.

Dococsahexaenoic acid is the reason for the low rate of caveolar transcytosis. Here is why.

[ Nature vol. 509 pp. 432 – 433, 503 – 506, 507 – 511 ’14 Neuron vol. 82 pp. 728 – 730 ’14 ] An orphan transporter, MFSD2a (Major Facilitator Superfamily Domain containing 2a) is selectively expressed in the BBB endothelium. It is REQUIRED for formation and maintenance of BBB integrity. Animals lacking MFSD2a show uninhibited bulk transcytosis across the endothelium. The animals show no obvious defects in the junctions between the endothelial cells. Pericytes (cells in the brain layer after the endothelium) are important in keeping the levels of MFSD2a at normal levels as animals lacking them show the same defects in the BBB as those lacking MFSD2a. Even though knockouts don’t have much of a BBB, they have normal patterning of vascular networks.

MFSD2a is the major transporter of docohexaenoic acid (DHA), an omega3 fatty acid (more later). DHA isn’t made in the brain and must be transported into it. Knockouts have reduced levels of DHA in the brain accompanied by neuronal loss in the hippocampus and cerebellum and microcephaly. Human cases due to mutation are now known (11/15). Transport of DHA and fatty acids into the brain across the BBB occurs only in the form of esters with lysophosphatidylcholines (LPCs) but not as free fatty acids in a sodium dependent manner. The phospho-zwitterionic headgroup of of LPC is essential for transport. MFSD2a ‘prefers’ long chain fatty acids (oleic, palmitic), failing to transport fatty acids with chain lengths under 14.

So MFSD2a inhibits transcytosis at the same time it promotes fatty acid transport into the brain. Major Facilitator Superfamily (MFS) proteins use the electrochemical potential of the cell to transport substrates. The best known MFSs are the glucose transporters (GLUT1 – 4).

So the blood brain barrier is due in part to the lipid transport activity of MFSD2a which gives BBB endothelium a different lipid composition (with lots of docosahexenoic acid) ) than others, inhibiting caveolar transport. Increased DHA levels are associated with membrane cholesterol depletion, as well as displacement of caveolin1 (the major protein involved in this form of transcytosis) from caveolae.

It is likely that MFSD2A acts as a lipid flippase, transporting phospholipids, including DHA containing species from the outer to the inner plasma membrane leaflet (where caveolin1 binds).

What is so hot about docosahexenoic acid — 22 carbons all in a row, a carboxyl group and 6 double bonds. We’re not talking fused ring systems, alkaloids, bizarre functional groups etc. etc.

Half the answer is that the double bonds are NOT randomly arranged. The 6 occur all in a row (but with methylene groups between them). This tells the chemist that they are not conjugated, hence the chain is probably not straight. Think how unlikely the arrangement is considering the way 6 double bonds and 9 methylenes COULD be arranged in a chain (2^15). Answer 5 ways depending on where the arrangement starts relative to the end of the chain.

The other half is that all the double bonds are cis, making it very unlikely that the 21 carbon chain can straighten out and cross the membrane. Lots of DHA means a very disordered membrane, which may be impossible to caveolin1 (and clathrin) to bind to.

So even though it’s years and years since I left organic chemistry, it permits the enjoying of the biochemical esthetics of the blood brain barrier.

The tight junctions between endothelial cells are primarily responsible for barrier function. These tight junctions are found only in the capillaries and postcapillary venules of the brain. Endothelial cells of the brain have few pinocytotic vesicles and fenestriae. [ Neuron vol. 71 p. 408 ’11 ] The brain vasculature has the thinnest endothelial cells, with the tightest junction and a higher degree of pericyte coverage coverage (‘up to’ 30%). [ Neuron vol. 78 pp. 214 – 232 ’13 ] The tight junctions are made from occludin, claudins and junctional adhesion molecules, and are closer to the lumen than the adherens junctions (which also link endothelial cells to each other) made by the cadherins (E, P and N). (ibid p. 219) TLR2/6 specific stimuli.