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placed by succinate, in which case photoreduction of DPN proceeds several times faster than in the presence of F M N H z ; here again T P N cannot replace DPN.
Vol. 80
THE REACTION OF PHOSPHINE METHYLENES WITH BORON HYDRIDES
Sir: Triphenylphosphine methylene (I, (CsHs)3PfCH2-) may be regarded as a carbanion stabilized by T TABLE I bonding of the unshared pair with adjacent phosRATESAND REQUIREMENTS OF LIGHTINDUCED OXIDATION- phorus orbitals. It therefore appeared probable REDUCTION REACTIONS that this material would interact with electron(Rates in pmoles per hr. per pmole chlorophylla) deficient boron hydrides such as diborane and deAll preparations made up to 3.0 ml. in 0.1M glycylglycine caborane to produce compounds containing P-C-I3 (pH 7.5) (chlorophyll content of chromatophores from 0.02 bonding. to 0.025 pM.). Further additions as indicated: DPN, Diborane Reactions.-Addition of gaseous diTPIV 0.7 p M . DPNH, 0.1 pM. FMIV, 0.15 p M . FMSHz (catalytically reduced with Hz), 0.05-0.15 pM. Samples borane to an ethereal solution of I in diethyl ether degassed in anaerobic cuvettes. a t room temperature resulted in rapid decolorizaDark period following tion and the deposition of triphenylphosphine -Light-illuminationSubs. Re- OxiSulis. Re0 methylene boron trihydride (11, (CsHs)aP+CHzAdditions meas. duced dized mens. duper1 dizel BHa-). The product, 11, proved to be stable 1 FMN FMN 0.0 toward water and was recrystallized easily from 2 FMNHz FMiYH2 . . . 0 . 3 FM.INH? . . . 0 . 3 methylene chloride-diethyl ether solution. Hy3 DPN + drolysis with aqueous hydrobromic or hydroDPNH DPNH . . . 0 . 2 DPSH . . . 0.2 chloric acids afforded three moles of hydrogen per DPX 9 . 1 . . . DPXH . . 0.6 4 DPN + mole of boron trihydride and the corresponding FMNHz FMNHz . . . 9 . 5 FMS 0.6 . . . triphenylmethylphosphonium halide. 5 4(+ D P S H DPNH . . . 0 . 6 DPYH . . . 0 . 6 ,
+
FMN FMN 0 . 6 . . . FMN 0.6 . . . formed in the light) heated 2 min. at 60" 6 TPN TPhT 0.0 FMKHz FMSHz . . . 0 . 3 FMKH? , . , 0 . 3 a Concentration changes measured a t 15-20' with a Reckman DU Spectrophotometer. Actual changes in optical density a t 340 mp of the order of 0.1 zt 0.005 unit for an initial 10 min. period of illumination (initial o.d. 0.5-0.8); dark values of the order of 0.01 j=0.002 unit per hour. FMK measured a t 455 nip, Difference spectra kindly measured by Dr. V. Lorber.
+
C B H ~ 'CH3BHaSP
+ 4HX
Hz0 ------f
+
(CGH~)BP+CHBX-3H2
+ BX?
Iodine was reduced to iodide ion and silver ion was reduced to silver metal by I1 in alcoholic solution. Table I presents the preparation data and properties of I1 and two other triphenylphosphine methylene boron trihydrides which have substituents on the methylene carbon. Each of these compounds displayed strong B-H stretching bands a t 4.40 and 4 . 5 0 ~ . TABLE I PROPERTIES O F TRIPHESYLPHOSPHINEMETHYLENE BOROSTRIHVDRIDES
The purified chromatophores, without any added enzymes, can carry out either photophosphorylations or photoreduction of DPN or both reactions simultaneously; photoreduction of DPN is partially inhibited when photophosphorylation occurs a t the same time. Vernon6 has described a system from R. rubrum, fortified with a number of enzymes, which preferentially photoreduces T P N (as indicated by suitable trapping agents); it is hoped that the basis for this difference in pyridine nucleotide specificity can be ascertained soon. The pyridine nucleotide specificity and the high lability of the system make i t plausible that one is dealing with an enzymatic reaction and not simply with a non-enzymatic, chlorophyll sensitized reaction.' Thus, the simultaneous stoichiometric reduction of DPN and oxidation of FMNHz in the light represent a reaction in bacterial preparations analogous to the Hill reaction of illuminated chloroplasts1t2(cf. 8).
PREP.4R.4TION
AND
A c a d . Sci. U S . S . R . ,6 7 , 32.5 (1949). ( 8 ) L. P . Vernon and hZ. D. Kamen, A r c h . Aiochem. Biobhys., S1,
(1) E. LViberg and P. Strebel. A n i $ . ,607, Y (1937). ( 2 ) Obtained from American Potash and Chemical Co , Henderson, Pievada.
R in (CsHs)rP*CHRBHa
hl.p., OC.
H CHB CbHs
191-192' 171-1 72" 143-li-L0 C
H CH3 CeHj
Vo Yield from Moles Hz (CsHa)aP+CHzRBr M o l e Compd.
Calculated, 5% H B
78.65 6.95 3.72 78.97 7 . 2 9 3.56 81.98 6 . 6 1 2.93
45 38 42
3 02
2 98 2 90 C
F o u nH d . "0
B
78.40 7.16 3.50 78.80 7 . 2 1 3.71 81.62 6 . 6 8 3 . 1 8
The formation of I1 from I and diborane is in sharp contrast to the results of Wiberg and Strebell who reported the reaction of ethylmagnesium halides and diborane to produce triethylborane and magnesium halohydrides, HXgX. Decaborane Reactions.-The addition of an ethereal solution of decaborane2 to a similar solution of I produced an oil which crystallized 011 standing to give a 35y0 yield of bright yellow DEPARTMENT OF BOTANY rhombs (111). The product was recrystallized easily VSIVERSITY OF MINSESOTA ALBERTU'. FRESKEL from methylene chloride-diethyl ether. m.p. 127hIINSEAPOLIS 14, MINNESOTA 129'; C19HelBloP(found: C, 56.8; H , S.01: I), RECEIVED M A Y 8, 1958 25.9; P, 7.GO. Calculated: C, 57.23; 11, 7.S4; ( 6 ) L. P . Vernon. THISJ O U R N A L , 80, 2.46 (1858); F e d e r a t i o ~Proc., B, 27.11; P, 7 . 7 ) . The infrared spectrum of I11 17,3 2 8 (19B8). contained B-H stretching a t 4 . 0 5 ~(terminal) and ( 7 ) A. A. Krasnovskii and G. P. Brin, Compl. Rend. ( D o k l n d y ) 1 2 2 (185.4).
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July 5, 1958
5 . 3 0 ~(bridge) as well as the bands commonly associated with the triphenylmethyl phosphonium ion. Acidification of I11 with dilute hydrochloric acid produced immediate decolorization and decaborane was recovered in 5oY0 yield. Air oxidation and hydrolysis of I11 was not apparent after three months a t ambient temperature. I t appears that this material is the triphenylmethylphosphonium salt of decaborane and is the first such material to be isolated as a pure stable (C6Hj)aP'CHz-
+ BioHir
---f
(C6Ha)3PtCHsBloH13-
substance. The reactions of other phosphine methylenes with decaborane are under investigation.
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when chromatographed with butanol-acetic acidwater,8 phenol-water,8 or pyridine-ethyl acetatewater.g I n addition, an ultraviolet absorbing compound was formed which exhibited the same chromatographic and spectral properties as guanosine diphosphate. Longer hydrolysis led to the formation of a second ultraviolet absorbing conipouiid which exhibited the same properties as guanosine monophosphate. I n view of the well-established role of the uridine sugar nucleotides as glycosyl donors in the biosynthesis of many complex saccharides, i t is an interesting variation to find fucose occurring in a guanosine nucleotide. The only other guanosine sugar nucleotide known a t present is guanosine diphosphate mannose.'O- l1
ROHM& HAASCOMPANY ARSENALRESEARCH DIVISION REDSTONE ALABAMA M. FREDERICK HAWTHORNE (8) S. M. Partridge, Biochem. J . , 42, 238 (1948). HUNTSVILLE, (9) M. A. Jermyn and F. A. Isherwood, ibid., 44, 402 (1949). RECEIVED MAY23, 1958 (IO) E. Cabih and L. P.Leloir, J . B i d . Chern., 206, 779 (1954).
Acta, 17, 283 (1965). (12) U. S. Public Health Service Postdoctoral Fellow. (11) J. L. Strominger, Biochim. e l Biophrs.
ISOLATION OF GUANOSINE DIPHOSPHATE FUCOSE FROM AEROBACTER AEROGENES
NATIONAL INSTITUTE OF ARTHRITIS& METABOLIC DISEASES INSTITUTES OF HEALTH NATIONAL
Sir: PUBLICHEALTHSERVICE V. GINSBURG UNITEDSTATES 14, MARYLAND H . N. KIRK MAN^^ We wish to report the isolation of a new sugar BETHESDA RECEIVED APRIL 5, 1958 nucleotide, guanosine diphosphate fucose, from a strain of Aerobacter aerogenesl that produces a polysaccharide containing L-fucose.2 ORGANOBORON COMPOUNDS. X. MIXED The nucleotides from 150 g. wet weight of bacTRIALKYLBORANES DISTILLABLE WITHOUT DISPROPORTIONATIONIJ teria were extracted with boiling 70% ethanol, precipitated with mercuric ion, and chromato- Sir: graphed on a Dowex 1-C1- c ~ l u m n . ~ Elution with It was suggested recently' that mixed trialkyl0.01 N HC1 and increasing concentrations of NaCl boranes characterized by the presence of a t-butyl yielded an ultraviolet absorbing peak which was group may manifest unusual stability to dispropordetermined to be 75% uridine diphosphate glucose tionation. One such substance, diisobutyl-t-butyland uridine diphosphate galactose by enzymatic borane, was described previously and its stability a n a l y ~ i s . ~This peak contained a total of 10 pM. to disproportionation was attributed to steric interof nucleotides calculated as uridine from spectral ference with the disproportionation mechanism. data. A minor component of this fraction amountWe wish to describe now the first distillable ing to 10% of its optical density a t 260 mp could be trialkylborane containing three dissimilar alkyl isolated b y paper electrophoresis or paper chro- groups, namely, t-butyl-isobutyl-n-amylborane. matography. The ultraviolet absorption spectrum This substance, b.p. 43.544.0" a t 0.5 mm., n2'D of this component was typical of a guanosine de- 1.4296, d Z 50.7506, was fractionally distilled twice rivative. I t s mobility during electrophoresis in in vacuo without decomposition, rearrangement or sodium formate buffer, p H 3.5, was less than guano- disproportionation. Anal. Calcd. for CI3H29B: sine diphosphate, but greater than guanosine B, 5.52. Found: B, 5.56. MRD: c a l ~ d . 67.30; ,~ monophosphate while it had a higher Rt than obsd., 67.48. Oxidation with alkaline hydrogen guanosine monophosphate when chromatographed peroxide' produced equimolar quantities of t-butyl, with ethanol-neutral ammonium acetate s o l ~ t i o n . ~isobutyl and n-amyl alcohols in high yield. The isolated component was analyzed colort-Butyl-isobutyl-n-amylborane was prepared in imetricallyo and found to contain approximately two ways: (a) in 50% yield by the alkylation of 0.8 pM. of 6-deoxyhexose per pM. guanosine. The n-amyldifluoroborane with t-butylmagnesium chloabsorption peak a t 400 mp given by the guanosine ride in anhydrous ether; (b) in 3Oy0 yield by the derivative in this test was identical in shape with reaction of t-butyl-di-n-amylborane with isobutylthat given by authentic fucose and also disap- magnesium bromide. The physical constants and peared a t the same rate upon dilution with water.I the infrared spectra of the two samples were pracHydrolysis of the guanosine derivative in 0.01 tically identical. I n connection with method (a), N HCI at 100" for 10 minutes liberated a com- i t is noteworthy that one t-butyl group derived pound having an Rr identical with that of fucose from the Grignard reagent rearranges to isobutyl (1) Strain AsSi (ATCC 12057). during the alkylation reaction. Concerning (2) J. F. Wilkinson, W. F. Dudman and G. 0. Aspinail, Biochem. J., 59, 446 (1955). (3) E . Cabib, L. F. Leloir and C. E. Cardini, J . B i d . Chem., 803, 1055 (1953). (4) H. M . Kalckar, E. P. Anderson, and K.J. Isselbacher, Biockim. Biophys. Acfa,20, 262 (1956). (5) A. C. Paladini and L. P.Leloir, Biochem. J . , 51,426 (1952). (6) 2. Dische and L. B. Shettles, J . B i d . Chem., 176, 595 (1918). (7) 2. Dische and L. B. Shettles, ibid., 192, 579 (1951).
(1) Previous paper, G. F. Hennion, P. A. McCusker and A. J. Rutkowski, THISJOURNAL, 80, 617 (1958). (2) Contribution from the Radiation Project operated by the University of Notre Dame and supported in part under Atomic Energy Commission Contract AT-(1 1)-38. (3) The B-C bond refraction was taken a s 1.93. (4) S . L. Clark and J. R . Jones, Abstracts, 133rd Meeting, American Chemical Society, San Francisco, April, 1958, p. 34-L.
