Nuclein and Nucleic Acids
All parts of an organism are essential for life. Only with this in mind does it make sense to say that the most important part of the cell is its nucleus. From the nuclei of cells in pus and in salmon sperm, Johann Friedrich Miescher (1811-1887) obtained a peculiar kind of substance, which he named nuclein (1868). Its phosphate content was easily discovered, but to find the exact proportions and the nature of the other components required special methods of separation from phosphatides and other proteins. It was difficult to develop such methods at a time when little was known about the properties, and particularly the stability, of a nuclein. For preparing nuclein from yeast cells, Felix Hoppe-Seyler (1825-1895) described the following details: Yeast is dispersed in water to extract soluble materials, like salts or sugars. After a few hours, the insoluble material is separated, washed once more with water, and then extracted with a very dilute solution of sodium hydroxide. The slightly alkaline solution, freed from insoluble residues, is slowly added to a weak hydrochloric acid. A precipitate forms which is separated by filtration, washed with dilute acid, then with cold alcohol, and finally extracted by boiling alcohol. The dried residue is the nuclein.32 It contains six percent phosphorus. A little more washing with water, a slightly longer treatment with acid or alcohol gives products of lower phosphorus content. Many experimental variations were necessary to establish the procedure that leads to purification without alteration of the natural substance.
This was also true for the methods of chemical degradation, carried out in order to find the components of nucleins in their highest state of natural complexity. It was learned for example, that the special kind of carbohydrate present in nucleins was very susceptible to change under the conditions of hydrolysis by acids. Phoebus Aaron Theodor Levine (1869-1940), therefore, used the digestion by a living organism. With E. S. London, he introduced a solution of nucleic acid into, e.g., the gastrointestinal segment of a dog through a gastric fistula and withdrew the product of digestion through an intestinal fistula. Fortunately, the products obtained in such degradations were not new in themselves. The carbohydrate in this nucleic acid proved to be identical with D-ribose, which Emil Fischer had artificially made from arabinose and named ribose to indicate this relationship (1891). The nitrogenous products of the degradation were identical with substances previously prepared in the long study of uric acid. In the course of this study, Emil Fischer established uric acid and a number of its derivatives as having the elementary skeleton of what he called “pure uric acid,” abbreviated to purine. Out of Adolf Baeyer’s work on barbituric acid came the knowledge of pyrimidine and its derivatives.
From these findings, together with what Oswald Schmiedeberg (1838-1921) had established concerning the presence of four phosphate groups in the molecule (1899), Robert Feulgen (1884-1955) constructed the following scheme of a nucleic acid. Feulgen’s formula of 1918 is:
Phosphoric acid—Carbohydrate—Guanine
Phosphoric acid—Carbohydrate—Cytosine
Phosphoric acid—Carbohydrate—Thymine
Phosphoric acid—Carbohydrate—Adenine
Of the four basic components on the right, thymine occurs in the nucleic acid from the thymus gland. Yeast contains uracil instead. The difference between these two bases is one methyl group: thymine is a 5-methyluracil. In all of these basic substances, the structure of urea
NH2 /
C=O
\
NH2
is involved, and they form pairs of oxidized and reduced states:
| Purine | Pyrimidine | |
| (reduced) Adenine | + | (oxidized) Thymine |
| (oxidized) Guanine | + | (reduced) Cytosine |
The exact position of phosphoric acid was established after long work and verified by synthesis.33
A compound of adenine, ribose, and phosphoric acid was found in yeast, blood, and in skeletal muscle of mammals. From 100 grams of such muscle, 0.35-0.40 grams of this compound were isolated. If the muscle is at rest, the compound contains three molecules of phosphoric acid, linked through oxygen atoms. It was named adenosine triphosphate or adenyltriphosphoric acid,34 usually abbreviated by the symbol ATP. It releases one phosphoric acid group very easily and goes over in the diphosphate, ADP, but it can also lose 2 P-groups as pyrophosphoric acid and leave the monophosphate, AMP.
This change of ATP was considered to be the main source of energy in muscle contraction by Otto Meyerhof.35 The corresponding derivatives of guanine, cytosine, and uracil were also found, and they are active in the temporary transfer of phosphoric acid groups in biological processes.
Thus, the study of organic phosphates progressed from the comparatively simple esters connected with fatty substances of organisms to the proteins and the nuclear substances of the cell. The proportional amount of phosphorus in the former was larger than in the latter; the actual importance and function in the life of organisms, however, is not measured by the quantity but determined by the special nature of the compounds.
The study of this function is the newest phase in the history of phosphorus and represents the culmination of the previous efforts. This newest phase developed out of an accidental discovery concerning one of the oldest organic-chemical industries, the production of alcohol by the fermentative action of yeast on sugar. A transition of carbohydrates through phosphate compounds to the end products of the fermentation process was found, and it gradually proved to be a kind of model for a host of biological processes.
Specific phosphates were thus found to be indispensable for life. In reverse, the wrong kind of phosphates can destroy life. As a result, an important part of the new phase in phosphorus history consisted in the study—and use—of antibiotic phosphorus compounds.