COMMUNICATIONS TO TIIE EDITOR
227s
VOI. 76
In a similar manner we have derived the following equation for the ratio (Cl4O2from 6-C14-glucose,/ C1402from l-C14-glucose),which we shall designate W
electrophoresis in 0.01 ;1.f Na2HP04,pH 8.8, a t I .-I3. Moving boundary electrophoresis in cacodylate buffer of ionic strength 0.1 and p H 6.5 a t 1.4' showed a component with the mobility of hemoglobin F and a component with a mobility greater (E'6R) E ) , whence = ( E / 6 R ) (1 than that of hemoglobin A but slightly less than 6RW that of sickle cell (S) hemoglobin. The mobilities E = 1 + W ( 6 R - 1) of hemoglobins A, S and C in this buffer are, reset.-' Again, using the experimental values of Bloom, spectively, 2.4, 2.9 and 3.2 X 10-5 et a1.,2.afor R and W , namely, 2 and 0.3, respec- volt-'.' The component having the mobility of tively, we calculate that 84YGof the COZ derived from hemoglobin F comprised 41% of the total by elecglucose was formed glycolytically, a value in good trophoretic analysis. This component was isolated agreement with that obtained from our equation (1). from the ascending limb of a moving boundary In our experiments with rat liver slices, the ob- experiment in 0.01 M Na2HP04; its ultraviolet served ratio U = a / b was higher than that reported absorption spectrum was found to be that of hemoby Bloom, et a1.,2 and varied between 1 and 1.8. globin F. The ultraviolet spectrum of the original However, these workers added lactate, acetate specimen did not differ significantly from that of a and gluconate, in high concentrations, to their comparable mixture of hemoglobins A and F." media. Under such conditions, as shown in Table When the original specimen was partially deI, Cl4OZformation from glucose is depressed, and natured with dilute sodium hydroxide, the amount the relative importance of the hexose monophos- of alkali-resistant hemoglobin recovered corresponded to the amount of the fetal electrophoretic phate shunt is increased. component in the specimen. The alkali-resistant TABLE I component was found to have the ultraviolet 250-mg. rat liver slices incubated with 2.5 ml. of Krehsspectrum of hemoglobin F. The non-fetal cornRinger bicarbonate buffer containing 55 pmoles of labeled ponent, isolated from the ascending limb of the glucose. The addition was 50 mm. each of lactate, acetate 5% Cor; incubated p H 6.5 moving boundary experiment, had the color and gluconate. Gas phase 95% 0 2 and visible spectrum of hemoglobin. The solufor 3 hours a t 37". bility of the original specimen as amorphous ferroLabel in yo of C" Ea X glucose Addition in C'4O2 L: = a / b 100mo hemoglobin in 2.58 M phosphate buffer of pH 6.8 even-C'd None a 2.8 a t 25' was 1.94 g. per liter, a value similar to those 1.2 94 of mixtures of hemoglobins A and F under the same 1-ci4 None b 2.8 condition^.^ We conclude from these results that the hemoglobin specimen examined consisted of a a 0.8 e ~ e n - C ~ ~+ mixture of hemoglobin F and a hitherto undescribed 0.6 79 abnormal human hemoglobin, which we shall call b 1.4 1-cl4 These values of E are calculated from our equation (1) hemoglobin E. It differs from all of the previously using the value of R = 2 determined experimentally hy described forms in its electrophoretic behavior. Bloom, et ~ 1 . ~ Its absorption spectrum, solubility and lability to Thus, in rat liver slices, over 90YG of the COZ alkali denaturation are similar to those of normal is derived from glucose via glycolysis, and even adult hemoglobin. A detailed account of this under the special conditions of Bloom, et aZ.,213 work will be published later. about SOY0 of the CO, is formed glycolytically. (3) G. H. Beaven, H. Hoch a n d E . R. Holiday, Biochem. J . , 49, 374
+
+
+
(1951).
DEPARTMENT OF PHYSIOLOGY J. KATZ (4) H. A . Itano, Arch. Biochem. B i n p h y s . , 47, 148 (1953). UNIVERSITY OF CALIFORNIA S. ABRAHAM OF CHEMISTRY SCHOOL OF MEDICINE R. HILL GATESAND CRELLINLABORATORIES No. 1889) BERKELEY, CALIFORNIA I. L. CHAIKOFF (COSTRIBUTION INSTITUTE OF TECHNOLOGY CALIFORNIA RECEIVED JANUARY 25, 1954 PASADEXA 4, CALIFORNIA -_ AND THE HARVEY A . ITAKO CHILDREN'S HOSPITAL, OF BIOCHEMISTRY AND OF PEDIATRICS DEPARTMENTS IDENTIFICATION OF A FOURTH ABNORMAL HUMAN USIVERSITY OF SOUTHERN CALIFORXIA IT.R . BERGREN HEMOGLOBIN LOSASGELES,CALIFORNIA PHILLIPSTURGEON Sir: RECEIVED MARCH22, 1951
I n addition to normal adult (A) and fetal (F) hemoglobins three abnormal forms (S, C and D) of human hemoglobin have been described. 1 , 2 We wish to report the identification of a fourth abnormal form in the erythrocytes of a child (M. &with I.)an atypical anemia. Filter paper electrophoresis of hemoglobin from this individual in 0.01 k?sodium barbital, PH 9.2, a t room temperature revealed two components, one with the mobility of hemoglobin F and the other with a mobility very nearly that of hemoglobin C. The same result was obtained by moving boundary II. A . Itano, S c i e n r e , 117,89 (1953). ( 2 ) J. V. Neel, et at., i b i d . , 118, 116 (1853). (1)
REACTION OF BIS-(CYCLOPENTAD1ENYL)-TITANIUM DICHLORIDE WITH ARYLLITHIUM COMPOUNDS'
Sir: Ris-(cyclopentadieny1)-iron was first described? in 1051. Since then, analogous complexes of n number of other transition elements have been ~ r e p a r e d . ~The structure of the iron complex has ( 1 ) This work was carried o i i t under Contract Nonr-R82(00) with t h e Office of Naval Research. ( 2 ) T. J. Kealy a n d P. L. Paiison. .\'aluve, 168, 1039 (19.51). (3) (a) G . Wilkinson, THISJ O U R N A L , 7 4 , 6146, 6148 (1952); 76, 209 (1954); (b) G. Wilkinson, P. L. Pauson, J . hl. Birmingham and F . A. Cotton, i b i d . , 76, 1011 (1953).
