Tuesday, March 9, 2021

Project GRFT: The story so far

In this and the posts to follow, we will introduce project GRFT, what this project is doing, and why it's important.  GRFT = Griffithsin.  The first part of our story starts here. 

**********************************************************************************************************************************************************************

Seaweed

In the 1800’s a widow, Mrs. Amelia Griffiths (1768-1858), roamed the seacoasts of Devon, Cornwall and Dorset with her wicker basket in hand, passionately collecting and cataloging the marine algae commonly known as seaweed.  In honor of Mrs. Griffiths and her contributions to the science of phycology, the Swedish marine botanist Carl Adolph Agardh named a genus of red marine algae “Griffithsia”. 

Griffithsia setacea from Griffiths’ books. © 2014 Royal Albert Memorial Museum & Art Gallery, Exeter City Council

Griffithsia are eukaryotic organisms of the Rhodophylum (Rhodo, from the ancient Greek, Rhodon, Rose and phylum which in the taxonomy stack is below kingdom and above class). Griffithsia are found in the oceans of the world. They dance gracefully in Neptune’s realm off the Coast of New Zealand. 

Viruses

In the late 1800’s, tobacco plantations in the Ukraine and Bessarabia (now Moldova) were, like many other plantations worldwide, infected with a disease that caused spots on the leaves causing a mottled appearance or mosaic pattern.  This was called mosaic disease because the patterns sometimes resembled mosaics.  The disease could and did destroy entire tobacco crops.  Scientists in Russia and Europe started an inquiry into the cause of the disease.  Was it a mold or a bacteria?  Was it something else?  Conducting experiments, they found that the agent which infected the plants passed through filters with a small pore size that would have stopped bacterial cells.  It could not be seen with the microscopes in use at the time.  It would not grow on prepared media on which bacteria or fungi would thrive.  So this was something as yet undiscovered.  Dmitry I. Ivanovsky and Martinus W. Beijerinck started the groundwork for what would become Virology, the study of viruses. 

What is a virus?  First we should look at what a virus is not.  Animal, vegetable, or mineral it is not.  Fungus or bacteria it is not.  Viruses can't be considered cells or independently living organisms. Perhaps viruses are best imagined as being exquisite alien machines running a program to inject nucleotides into  living cells causing the cellular mechanism to produce more of the virus.  Viruses are made of nucleotides often dressed up in a protein coat or capsid. 

Not all viruses cause health problems.  Some we find useful for a wide variety of applications including Biomanufacturing by the expression of of recombinant protein in organisms.  Viruses considered pathogenic to humans only number about 100 - 200 varieties. These disease causing viruses, however, represent a major health concern and are a significant cause of mortality and suffering worldwide.

Prevention of viral outbreaks

 

Prevention of the spread of pathogenic viruses may be accomplished in several ways, for example: 

1.        Physical distancing, physical barriers.

These keep the virus particles, known as virions, away from cells by methods such as masking and wearing other PPE, also by air filtration.

 

2.       Vaccines

Vaccination stimulates the immune response with the body producing antibodies against the particular virus.   Vaccination usually only requires 1-3 doses of the vaccine and may provide immunization for a lifetime.

 

 

Antivirals

Antivirals prevent the virus from hijacking cells in the host and replicating.  The antiviral works typically by blocking entry into the cell by the virus.  Antivirals need to be taken whenever there is danger of the transmission of a virus.  Antivirals generally work against pathogenic viruses when administered before or at the first stages of infection.  


Antiviral lectins

A lectin may be defined as a carbohydrate binding protein. 

Lectins are found in everywhere in nature including in the food we eat. 

An example of an antiviral lectin is BanLec or banana lectin.  Found in the bananas Musa acuminata and Musa balbisiana, BanLec has been shown to be effective against HIV. 

Other examples of effective antiviral lectins found in nature are cyanovirin-N, scytovirin, and microvirin.   

Griffithsin

   Of all the known antiviral lectins found in nature, perhaps griffithsin, the red seaweed protein, is the shining star. Besides being free of significant toxicity, griffithsin is the most powerful of any of the lectins at inhibiting enveloped viruses from entering cells.  

Griffithsin, or GRFT is a 121 amino acid, 12.7 kDa protein.



Wild type GRFT:

Please take a look at position 31 above.   In wild GRFT, this is a non-standard amino acid and in recombinant GRFT this is generally replaced by substituting alanine (the red A).

The yellow arrows designate the secondary B-sheet structures of the protein.

GRFT is a dimer and has 3 triangular prismatic blades in the Beta sheets.  These separately have been shown to have some antiviral properties, however together in griffithsin’s domain swapped dimer they are more powerful, just as two hands can grab and hold with more strength than one hand.  This dimeric structure has six carbohydrate binding sites that work together.  

An interactive 3D model of the GRFT protein structure may be found here, scroll down to "Structure" and play with it.

 

Future episodes to follow!

No comments:

Post a Comment