148 J. R. SIMMONS AND H. K. MITCHELL 
mistake to take this as evidence that all of the radioactive components are actually 
ninhydrin-positive. 
On hydrolysis all of the radioactive components yielded glutamic acid, though this 
was not always the only radioactive component found. All of the radioactive areas, 
Counts / min 
ine) 
oO 
Oo 
fe) 
fo) 
9 6 5 AEE 
Fig. 1. Distribution of radioactivity resulting from the paper chromatography of squashed whole 
animals injected with ['C] glutamic acid (4 wC/15 ul). Numbers indicate areas measured by plani- 
meter with 1 being the chromatographic origin and 3 most closely matching the position of known 
glutamic acid. 
when eluted and hydrolyzed, yielded more than one ninhydrin-positive spot following 
chromatographic separation. 
It was found that 4h of starvation prior to the injection of the {[14C] glutamic acid 
did not appreciably alter the pattern of radioactivity found following chromatog- 
raphy. Increasing the amount of radioactive amino acid injected 4-fold likewise 
failed to alter the pattern observed. 
Leucine incorporation. The results given in Table II and Fig. 2 are examples of 
those obtained in three experiments in which animals injected with leucine were 
squashed and chromatographed. As was the case with glutamic acid, the pattern of 
radioactivity resulting from the injection of leucine was very complex. It differs from 
DABBLE a 
GLUTAMIC ACID INCORPORATION 
84-h larvae were injected with a solution of 4 wC/15 wl of L-glutamic acid and three 
larvae were squashed on Whatman No. 1 paper at the time from injection indicated. 
After chromatography in the propanol-NH, solvent the chromatograms were 
scanned with the strip counter and the areas determined by planimetry. Radio- 
activity is reported as the per cent of total radioactivity present. Areas refer to those 
designated in Fig. 1. 




Time Total % of total radioactivity at area 
(min) area 5 D 3 4 5 6 
I 1.855 1.5 5-7 77.8 8.3 6.9 0.0 
5 1.955 2.5 Q.2 62.0 11.8 2a 2.4 
15 3.885 9.0 9.7 42.7 16.6 17.4 3.9 
30 .940 10.0 10.0 32.4 2351 16.8 7.8 
60 2.069 13.5 To? 30.0 22.6 22.0 4.7 

References p. 155 
