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Vitamin D Final Draft
Vitamin D Final Draft
D3 can isomerize to vitamin D3 within 2.5 hours in the skin. Within 1224 hours after
UVB exposure, the circulating concentrations of vitamin D3 are at their maximum levels.3
If previtamin D3 is formed in the skin, it can also undergo either photoisomerization to
lumisterol, tachysterol, and toxisterols, or it can be retransformed to 7-DHC.3 A broad
overview of this process can be seen in Figure 1.
At this point, the vitamin D is biologically inactive and must be activated in the
liver and the kidneys. The vitamin D found in vitamin supplements is also in
this inactive form. Inactive vitamin D is transported to the liver where it
is enzymatically hydroxylated to 25-hydroxyvitamin D [25(OH)D].5 The
hydroxylation into 25-hydroxyvitamin D
[25(OH)D] is catalyzed by microsomal cytochrome
P450 enzyme CYP2R1 and/or the mitochondrial
cytochrome P450 CYP27A1 (see Figure 2), both of
which are constitutively expressed.3,6 Cytochromes are
hemoproteins that supply ATP via electron transport to the
molecules. Besides the CYP2R1 and CYP27A1, there are
also
(CYP
between 25 nmol/L and 200 nmol/L.3 Currently, vitamin D levels of about 30 ng/mL
(75 nmol/L) are considered to be optimal for health.
25-hydroxyvitamin D [25(OH)D] is then transported to the kidneys and is finally
hydroxylated by CYP27B1 (25-hydroxyvitamin D-1-hydroxylase or 1OHase) at the
C1 position to hormonally active 1,25-dihydroxyvitamin D [1,25(OH)2D].3 The overall
activation of vitamin D3 can be seen in Figure 2 where there are two separate pathways
25(OH)D can take to form two different active forms of vitamin D or return to the
inactive form as 24,25(OH)2D.5 Calcitriol has biological effects on the kidneys but is
usually sent into the bloodstream to be used by other parts of the body.
Distribution of Vitamin D to the Rest of the Body
Once in the bloodstream, vitamin D in the form of active 1,25(OH)2D or
25(OH)D often binds to a protein called gc-globulin (group-specific component of
serum), also known as vitamin D binding protein (DBP)
(see Figure 3).7 This protein can be found in plasma and
cerebrospinal fluid, where it binds to vitamin D metabolites
and transports them to target organs. DBP belongs to the
Figure 3: Ribbon diagram of
vitamin D binding protein.21
Cells in the body have several ways of accessing the bodys supply of vitamin D
[1,25(OH)2D and 25(OH)D]. One way is by simple diffusion of free vitamin D across the
cell membrane. The level of free vitamin D in the body is governed by the levels and
affinity of DBP. Vitamin D can also enter the cell while still bound to DBP through
active-receptor-mediated uptake. The DBP-vitamin D complex binds to a cell surface
receptor called megalin (LRP2) and is internalized in a vesicle, where vitamin D is
released and DBP is denatured.7 Once inside cells, 1,25(OH)2D directs vitamin Ddependent gene regulation through the vitamin D receptor (VDR), while 25(OH)D is first
converted into active 1,25(OH)2D through hydroxylation of the 1- carbon by CYP27B1,
usually located in the inner mitochondrial membrane.7,8 This oxidation reaction of
25(OH)D occurs when NADPH-CYP reductase captures an electron pair from the
conversion of NADPH to NADP and transfers it to CYP27B1, which has high specificity
for 25-hydroxylated steroids (i.e. 25(OH)D) and reduces oxygen via the heme group in its
active site to hydroxylate 25(OH)D.6,8
The mechanism is likely similar to the
reduction of oxygen to water, which
involves heme D, the site of oxygen
reduction in many types of bacteria.20
Another cytochrome, CYP24A1,
Figure 4: Binding of 1,25D to CYP24A1 located in the
inner mitochondrial membrane. The heme group (red dotted
sphere) reduces oxygen to hydroxylate 1,25D.22
pressure, boost the immune system, or even help prevent cancer. Looking through the
literature, there are studies showing that vitamin D can alter dopamine, acetylcholine, and
noradrenaline neurotransmission; helps prevent onset of Parkinsons, schizophrenia,
depression, and other mental illnesses; and plays a multifaceted role in brain
development.9,11
define specific locations where vitamin D can initiate biological responses. Some of these
responses include the classic calcium homeostasis system, along with five other systems,
including the brain, which will be focused on later. The ligand-receptor complex
produces the biological reactions.
The first class of response that can be induced is the traditional genomic response.
VDR is a DNA-binding transcription factor consisting of a heterodimer (two different
molecules bound together, usually macromolecules in this case, the VDR with the
vitamin D ligand, as well as an unoccupied retinoid X receptor RXR see figure 6).13
After the ligand binds to the VDR genomic pocket (GP), there is a conformational change
to allow it to serve as a platform for coactivator binding. The coactivator allosterically
stabilizes the VDR-RXR heterodimer, allowing it to be phosphorylated by serine protein
kinases. This new complex can positively and negatively regulate gene transcription by
recognizing vitamin D response elements (VDREs) in DNA. The VDR-RXR then
recruits additional comodulators to help initiate transcription. There are many hypotheses
concerning how exactly this happens; Dr. Hausslers team proposes that there is a
simultaneous binding of multiple factors in a supercomplex at the promoter, based on the
model RANKL gene promoter. Activated VDR can also interact with transcriptional
coregulators to control gene expression. VDR consists of 427 amino acids with two main
functional groups: a zinc finger DNA binding domain near the N-terminus and a vitamin
D ligand binding domain near the C-terminus. A structure consisting of 12 -helices
allows VDR to heterodimerize with the retinoid X receptor.13
The practical implications come from finding the genes that are directly regulated
by this VDR complex. So far, at least eleven genes that encode bone and mineral
homeostasis (the traditional target of VDR) have been found, including gene products
that facilitate intestinal calcium intake. Another network that has been found to be
regulated by VDR is the encoding factors that impact cell life/cancer, the immune system,
and metabolism. These come from inducing and repressing various genes involved in
diseases such as type I diabetes, multiple sclerosis, and arthritis. It has even been found to
blunt various genes involved in inflammatory responses, thus reducing the risk of heart
disease and Alzheimers. It is clear that there are many areas regulated by VDR, and there
will certainly be more to come.
Vitamin D also plays a role in a second category of responses: rapid responses.
This cannot be explained by VDR-mediated gene transcription, as was shown in the
classical genomic responses. This is a relatively new area of research; the first rapid
responses were discovered in the 1980s from the rapid hormonal simulation of intestinal
calcium absorption in chicks. The transfer of calcium to the intestine was noticed only
after 4-5 minutes after transfer of vitamin D to the celiac artery. The main difference that
separates genomic and rapid responses is the time delay; genomic responses often take
days while the rapid response pathway takes
mere minutes. However, rapid responses are also often induced through a different
mechanistic pathway. The first clue came after it was noticed that only the 6-s-cis locked
and not the 6-s-trans locked analog was capable of producing rapid responses in the chick
model. This also means that the VDR also can adopt different conformers the so called
VDR-GP for genomic responses, the membrane caveolae localized VDR-AP for
alternative binding, and the plasma vitamin D-binding protein (DBP).
In particular, it has been found that the caveolae is the source of many rapidly
responding signal transduction pathways. Caveolae are located in the plasma membrane
and are enriched in sphingolipids and cholesterol. It was demonstrated both that vitamin
D showed the same binding affinities to VDR in the caveolae as observed with nuclear
VDR, and that vitamin D localized in vivo in the plasma membrane caveolae.14
Finally, the VDR AP site was proposed to resolve the VDR paradox (see Figure
7).23 Traditionally, only a
single ligand binding
domain has been
recognized the one that
binds only the 6-s-trans shape. However, the
rapid response
conformer is not able
Figure 7: The proposed VDS-VDR conformational ensemble model. The left panel
shows the conformational flexibility of VDS, the middle panel shows the different
binding sites on VDR, with the yellow oval showing overlap between the two regions.
The right panel shows specific conformational dynamics of VDR AF2 domain; the
Boltzmann distribution is altered depending on the nature of the ligand, changing the
energy landscape of VDR ensemble members to bias a specific downstream event.
to dock to this specific binding site. Computational work showed that there is an
alternative binding site available. This conformational model was proposed by Dr.
Haussler and his team, whereby the VDR could accommodate differently shaped ligands
to initiate both genomic and rapid responses. The steroid hormone would essentially test
the waters and form a receptor-hormone complex with the receptor species that provided
the highest affinity and most stability. Figure 8 shows the main differences between the
genomic and rapid response pathways.15 It seems as though the two categories are vastly
different, but a small portion of VDRs at the membrane is now believed to regulate the
expression of genes, thus regulating the activity of many kinases, phosphatases, and ion
channels. However, more research needs to be done to further elucidate the mechanisms
behind this process.
Implications of
Vitamin D/VDR
Figure 8: Different mechanisms by which vitamin D and VDR can induce chemical
responses in the body. On the left, the caveolae-related pathway leads to activation of the
secondary messenger system to elicit short term responses. On the right, 1,25(OH) 2D can
interact with VDR localized in the cell nucleus to produce genomic responses through
gene transcription.
and Sleep:
Since VDR has recently been found in the brain, some interesting new hypotheses
have emerged concerning the function of vitamin D in the brain.18 One of these concerns
the role of vitamin D in sleep. Normally, sleep is highly organized. Humans typically go
to sleep at the same time every night, going through specific phases, such as REM, slowwave, etc. Waking up from sleep is involuntary and also occurs in a fairly stable manner.
This seems to imply that sleep is not caused by a buildup of sleep-inducing hormones and
substances; rather, it is a result of a circadian rhythm type function where sleep is highly
coordinated by the time of day. This implies that one of the reasons for sleep problems
stems from brain chemistry.
Normally, many different hormones are secreted before sleep, leading to
drowsiness a warning, so to speak. Antidiuretic hormone is produced to limit urine
production, melatonin levels increase, and so on. The brain also induces paralysis as deep
sleep arrives, activating specific neurotransmitters to turn off the signals responsible for
wakefulness. These two categories, timing and paralysis, are essential to sleep. Saper and
colleagues suggest an on-off switch mechanism, which is responsible for sleep; one part
of the brainstem is active while the other is suppressed.16 Specifically, the hypothalamus
is thought to be involved because the stimulation of the posterior hypothalamus induces
arousal, while stimulation of the anterior hypothalamus and adjacent basal forebrain
region causes sleep.17 Now, how does this tie in with vitamin D? Vitamin D targeted
neurons have been discovered in specific brain and spinal cord locations in multiple
animals. This suggests a possible role of vitamin D in regulating sleep. A 2-year
uncontrolled trial of vitamin D supplementation in 1500 patients with neurological
complaints and sleep problems saw improvements in both these functions.18 Further
research needs to be done in this area, as sleep is also influenced by sociological and
psychological factors. For example, pain has been shown to influence the quality of sleep,
but pain has also been linked to vitamin D.19 Therefore, vitamin D may ameliorate the
quality of sleep through a multitude of factors not only through chemical pathways in
the brain, but also through alleviating pain. There is a delicate balance between these
factors; vitamin D could directly impact sleep, which could then improve feelings of
pain. Vitamin D could also improve a multitude of variables, including mood, quality of
life, etc. which could also improve pain (a subjective feeling that could be influenced by
psychology). Further research needs to be done to elucidate the function of vitamin D in
these processes.
Vitamin D has clearly grown in importance over the last half century. The
involvement of vitamin D in vital bodily processes, from bone health to even regulating
gene expression, reveals how potent this single hormone is to human health. New models
to elucidate the nature of vitamin D and VDR binding will further this cause and may
even reveal new routes for drug development and the curing of diseases, one of the most
fundamental concerns for the human race.
References
1. National Cancer Institute. "Vitamin D and Cancer Prevention." National Cancer
Institute. National Institutes of Health, n.d. Web. 29 May 2013.
2. Comparison of vitamin D2 and vitamin D3 supplementation in raising serum 25hydroxyvitamin D status: a systematic review and meta-analysis Laura
Tripkovic, Helen Lambert, Kathryn Hart, et al., The American Journal of Clinical
Nutrition 2012, 95, 1357-1364.
3. Vitamin D Metabolism Bodo Lehmann, and Michael Meurer, Dermatologic
Therapy 2010, 23, 2-12.
4. The Circadian Control of Skin and Cutaneous Photodamage Joshua A.
Desotelle, Melissa J. Wilking, and Ahmad Nihal, Photochemistry and
Photobiology 2012, 88, 1037-1047.
5. Vitamin D: Metabolism, Molecular Mechanisms, and Mutations to
Malignancies Natalie Nemazannikova, and Antonas Kiriakos Molecular
Carcinogenesis 2013,
6. Cytochromes P450 are essential players in the vitamin D signaling system Inge
Schuster, Biochimica et Biophysica Acta 2011, 1814, 186-199.
7. New perspectives on the vitamin D binding protein R.F. Chun, Cell
Biochemistry and Function 2012, 30, 445-456.
8. Enzymes involved in the activation and inactivation of vitamin D D.E.
Prosser, G Jones Trends In Biochemical Sciences 2004, 29, 664-667.
9. The effects of vitamin D on brain development and adult brain function
James P. Kesby, Darryl W. Eyles, Thomas H.J. Burne, et al., Molecular and
Cellular Endocrinology 2011, 347, 121-127.
10.
Distribution of the vitamin D receptor and 1 alpha-hydroxylase in human
brain D.W. Eyles, S. Smith, R. Kinobe et al. Journal Of Chemical
Neuroanatomy 2005, 29, 2130.
11.
Vitamin D, effects on brain development, adult brain function and the
links between low levels of vitamin D and neuropsychiatric disease D.W. Eyles,
T.H.J. Burnes, and J.J. McGrath, Frontiers of Neuroendocrinology 2013, 34, 4764.
12.
Haussler, Mark R., and Kerr Whitfield. "Molecular Mechanisms of
Vitamin D Action." Calcified Tissue International 92.2 (2013): 77-
Receptor Ensemble Model Offers Unique Insights into Both Genomic and
Rapid-Response Signaling." Sci. Signal 2.75 (2009): n. pag. Print.