COMMUNICATIONS TO THE EDITOR
Feb. 5, 1956
695
imino carbon from formiminoglycine (C, of purine) may become available for hydro~ymethylation,~ thus implying a variety of reactions and enzymatic steps beyond the postulated “formimino-tetrahydrofolic acid” intermediate. The activity of tetraTABLE I hydrofolic acid with these unfractionated extracts does not impinge upon the coenzyme form of this FUNCTION OF TETRAHYDROFOLIC ACID IN FORMIMINOGLYfactor. lo CINE DEGRADATION The accumulation of formiminoglutamic acid Thunberg tube, 4.5 ml.; 100 p M . phosphate buffer, pH 7.5; 3 p M . Na&04; extract (4 mg. bacterial nitrogen); from histidine in folic acid deficient rats,11.12and in 1 mg. THFA where indicated (prepared according to Bro- preparations of mammalian liverL3and of microquist, THISJOURSAL,73, 3535 (1951) half life in absence of organisms, l 4 suggests a similar sequence for this 0 2 about 2 weeks); after evacuation, 40 pM. substrate substrate and thus that formimino transfer may tipped from side arm; incubated 60’ a t 35’. constitute a biologically important reaction type, SubProducts
a stoichiometric yield of glycine and ammonia. N-Formylglycine was inactive and thus not considered as an intermediate in the conversion of formiminoglycine to glycine.
strate formed used, NHa fiM. Glycine fiM.
Substrate and enzyme
Formiminoglycine, 40 pM. Extract, crude Extract, Dowex Extract, Dowex THFA, 1 mg. N-Formylglycine, 40 pM. Extract, crude Extract, Dowex f THFA, 1 mg.
+
40 0 38
21 0 36
56 0 42
0 0
0 0
..
..
These observations suggested to us that formiminotetrahydrofolic acid might occur as an intermediate as Formiminoglycine
+ THFA
Formimino-THFA
glycine Hz0
+ formimino-THFA
LABORATORY OF BACTERIOLOGY R. D. SAGERS’~ BRIGHAMYOUNGUNIVERSITY, PROVO,UTAH J. V. BECK LABORATORIES OF BACTERIOLOGY AND CHEMISTRY W. GRUBER UNIVERSITY OF ILLINOIS,URBANA I. C. GUNSALUS RECEIVEDDECEMBER 28, 1955 (9) R. D.Sagers and J . B. Beck, J . Bect., in press. (10) B. E. Wright, THISJOURNAL, 77, 3930 (1955). (11) M.Silverman, R. C. Gardiner and H. A. Bakerman, J. Biol. Chem., 194, 815 (1952). (12) H. Tabor, M. Silverman, A. H. Mehler, F. S. Daft and H. Bauer, THISJOURNAL, 76, 755 (1953). (13) B . A. Borek and H. Waelsch, J. B i d . Chem., 205, 459 (1953). (14) H.Tabor and A. H. Mehler, ibid., 210,559 (1954). (15) Presently NSF Predoctoral Fellow, University of Illinois.
(2)
THE
STEREOCHEMISTRY OF BASE-CATALYZED ADDITIONS OF THIOLS TO ACETYLENES
Sir: THFA + ammonia + formate (3) To test this postulate, 40 ~LM.g l y ~ i n e - 2 - C ~ ~ Although a high degree of stereospecificity has and 40 pM. unlabeled formiminoglycine were in- been realized for the addition of hydrogen, halocubated with “Dowexed” extract, alone and with gens and hydrogen halides to acetylenes, similar added tetrahydro-folic acid. Aliquots were chro- studies on nucleophilic additions have not been matographed as described above and the chromato- published.‘ We have now observed that basegraphs scanned for radioactivity with an actigraph catalyzed additions of thiols to the acetylenic (Nuclear instrument, i210del C-100) previously compounds, phenylacetylene, 2-butyne, chlorocalibrated with g l ~ c i n e - 2 - Cof~ ~known activity. acetylene and P-tolylmercaptoacetylene, proceed As shown in Table 11, the radioactivity of glycine- in a trans fashion. Refluxing an alcoholic solution of phenylacetyl2-C14 was equilibrated rapidly with formiminoglycine in a tetrahydrofolic acid and enzyme de- ene with sodium p-toluenethiolate for 15 hours pendent reaction. Formylglycine does not ex- resulted in a 79% yield of cis-w-styryl p-tolyl change with glycine-2-C1* nor form formimino- sulfide (none of the trans isomer was isolated), glycine. Hydrolysis of the formiminoglycine which was readily oxidized by hydrogen peroxide formed yielded CL4in the glycine only. The re- to its sulfone, m.p. 76-77’, Xmax 266 mp, Emax action sequence beyond “formimino-tetrahydro- 14 X lo3; trans formj2 m.p. 121’, Xmax 276 mp, folic acid,” of which one route leads to ammonia Emax 25.9 X loa. trans-w-Styryl p-tolyl sulfone plus formate (reaction 3), is slower than the ex- also has been prepared by the Friedel-Crafts change. Preliminary data indicate that the form- reaction of trans-w-styrenesulfonyl chloride3 with toluene.‘ Similar results were obtained with sodium TABLE I1 methanethiolate and phenylacetylene, the product FORMIMINO TRANSFER TO GLYCINE-2-C14; TETRAHYDRObeing methyl cis-w-styryl sulfide, yield 73%, b.p. FOLIC ACID DEPENDENCE Protocol as Table I ; substrates, 40 pM. glycine-2-Cl4 101.5O (5 mm.). (Anal. Calcd. for C9HloS: C, 71.95; HI 6.71. Found: C, 71.28; H, 6.88.) (SA 3150 dis./sec./pM.) ; 40 p M . formiminoglycine; 1 mg. THFA where added. Its sulfone, m.p. 66-67’, Am, 261 mp, emax 19.9 C”, dis./sec./fiM. X lo3, was different from the trans isomer4, m.p. FormiminoIncubation, min. Glycine glycine 78-79’, Xmax 264 mp, emax23.7 X lo3. The 78-79’ 0 2 5 15 15, w/o extract 15, w/o THFA
3150 2210 1650 1530 3150 3150
0 905 1230 1310 0 0
(1) R. A. Raphael, “Acetylenic Compounds in Organic Synthesis,” Academic Press, Inc., New York, 1955,p. 43. (2) E.P. Kohler and H. A. Potter, THISJOURNAL, 57, 1316 (1935). (3) A. P. Terent’ev, R. A. Gracheva and Z . F. Shcherbatova, Doklady Akad. Nauk S.S.S.R., 8 4 , 975 (1952); C. S. Rondestvedt and J. C. Wygant, TEISJOURNAL, 7S, 5785 (1951). (4) W. E. Truce, J. A. Simms and H. E. Hill, ibid., 76, 5411 (1953).
BOOKSRECEIVED
Vol. 78
=-I ,R’ . This conm.p. isomer showed the characteristic trans absorption band a t 10.35 p . Both of these com- intermediate carbanion, R,C=C\R,t pounds were reduced to methyl b-phenylethyl figuration of the carbanion would be expected on sulfone. the basis that in the transition state the negative With 2-butyne, an alcohol solution of sodium charge (electron pair) and the negatively charged p-toluenethiolate reacted to give a 65Y0 yield of thiolate group would tend to be as far apart as 2-P-tolylmercapto-trans-2-butene, b.p. 83-88’ (2 possible. Considering the stereochemistry of the mm.), nZoD 1.5634. (Anal. Calcd. for CllHllS: isoelectronic oximes, such a carbanion would be C, 74.10; H, 7.91. Found: C, 73.82; H, 7.87). expected to be geometrically stable. However, This product has a strong infrared absorption band the mechanism of addition may also involve a a t 7.70 p ; trans-2-butene and 2-chloro-trans-2- synchronous process. Evidence is now being butene have a similar band a t 7.80 p which is absent sought to clarify this point. in their cis isomers. WILLIAME. TRUCE DEPARTMENT OF CHEMISTRY Chloroacetylene was treated with an alcoholic PURDUE JOHN A. SIMMS UNIVERSITY solution of an equimolar amount of sodium p- LAFAYETTE, INDIANA MAXM. BOUDAXIAN toluenethiolate to give [in addition to a 52.270 RECEIVED DECEMBER 17, 1955 conversion (77.4% yield) to cis-bis-(p-tolylmercapto)-ethene5] cis-l-chloro-2-(~-tolylmercapto)ethene (13.7% conversion, 20.3% yield), b.p. BOOKS RECEIVED 99-102’ (2.2 mm.), n Z o D 1.5901, and having a strong characteristic absorption a t 7.78 p . This December 10, 1955-January 10, 1956 band is absent in the spectrum of the trans isomer E. R . ANDREW. “Kuclear Magnetic Resonance.” Camas prepared by adding p-toluenesulfenyl chloride bridge University Press (American Branch), 32 E . 57th to acetylene6; neither of these isomers showed Street, New York 22, N. Y. 1956. 265 pp. $6.50. infrared absorption bands associated with the JAMES G. BECKERLEY,Editor, MARTIND. KAMENAND LEONARD I. SCHIFF,Associate Editors. “Annual Review group, >C=CHz. (Anal. Calcd. for the cis of Nuclear Science.” Volume 5. Annual Reviews, Inc., sulfone, m.p. 39-40’, C9H9SOzC1: C, 49.S4; H, Stanford, California. 1955. 448 pp. $7.00. 4.19; C1, 16.36. Found: C, 50.17; H , 4.80; R . BROWNAND J. F. DANIELLI(Editors). Symposia of,Fhe C1, 16.00). The rule of trans elimination’ [cis-1Society for Experimental Biology. riumber IX. Fichloro-2-(p-tolylmercapto)-ethene undergoes the brous Proteins and Their Biological Significance.” Acasodium ethoxide-promoted reaction with thiolate demic Press, Inc., Publishers, 125 East 23rd Street, New York 10, N. Y. 1955. 370pp. $8.00. reagent to form the cis-bis(ary1mercapto)-ethene much more readily than the trans isomer does] J. W. COOK(Editor). ”Progress in Organic Chemistry.” Volume 3. Academic Press, Inc., Publishers, 125 East also supports these assignments of configuration. 23rd Street, New York 10. N. Y . 1955. 273 pp. $7.80. p-Tolylmercap toacetylene, b .p . 73.O-77.0’ (2.5 mm.), Z ~ O D 1.5721. (Anal. Calcd. for mercuric J. FRENKEL. “Kinetic Theory of Liquids.” Dover Publications, Inc., 920 Broadway, hTewYork 10, N. Y . 1955. deriv., (CsH7S)zHg: C, 43.65; H, 2.85; S,12.96; 488 pp. $3.95 cloth, $1.95 paper. Hg, 40.52. Found: C, 43.50; H, 2.87; S,12.84; R “Methoden der Organischen Hg, 40.47), characteristic infrared absorption EUGENM ~ ~ L L E(Editor). Chemie.” (Houben-Weyl). Band IX. M. BOGEMANN, bands a t 3.08 and 4.90 p , on treatment with an H. BOHEE, H. ECKOLDT, J. GOERDELER, F. MUTH,S. alcoholic solution of an equimolar amount of PETERSEN,M. QUAEDVLIEG, H. RHEINBOLDT, A. SCHOBERL,A. SCHONBERG, 0.-E. SCHULTZ, H. SOLL A N D A . sodium p-toluenethiolate gave cis-bis-(p-tolylWAGNER. “Schwefel-, Selen-, Tellur-Verbindungen.” mercapto)-ethene5t8(77% conversion) without any Georg Thietne Verlag, Stuttgart, Germany. 1955. 1337 of its trans isomer being isolated. pp. DM 218.--. These nucleophilic additions may be proceeding KENNETHM. SMITHA N D MAX A. LAUFFER(edited by). by a stepwise mechanism, with formation of the “Advances in Virus Research.” Volume 111. Academic (5) W. E. Truce and R. J. McManimie, THISJOURNAL, 76, 5745 (1954). (6) W. E. Truce and hi. 31. Boudakian, unpublished results. (7) S. J. Cristol, Abstracts, 14th National Organic Chemistry Symposium, Am. Chem. SOC.,Lafayette, Ind., June, 1935. (8) W. E. Truce, M. M. Boudakian, R. F. Heine and R. J. McManimie, paper presented before the Division of Organic Chemistry, hmerican Chemical Society, September 1955, Minneapolis, Minnesota, Page 25-0.
Press, Inc., Publishers, 125 East 23rd Street, New York 10, N. Y. 1955. 339 pp. $8.00. WILLIAMS. SPECTOR (edited by). “Handbook of Toxicology.” Volume I. W. B. Saunders Company, Philadelphia, Pennsylvania. 1955. 408 pp. $6.75. ERICH VINCKE. “Darstellung von Hormonpraparaten.” S. Hirzel Verlag, Schuhmachergasschen 1-3, Leipzig C 1, Germany. 1955. 253 pp. Gzln, DM. 10.30.
