TY - JOUR
T1 - Serine hydroxymethyltransferase
T2 - Evidence for its presence in human, monkey and rat lenses
AU - Geller, Arthur M.
AU - Zigler, J. Samuel
AU - Jernigan, Howard M.
N1 - Funding Information:
Grosvenor of the UT, Memphis Department of Physiology for the rat pup lenses, Mr John P. Cogan, Chief, In Vitro Vaccine Testing Section, O&e of Biologics. National Center for Drugs and Biologics, for the monkey eyes, the Mid-South Eye Bank, Memphis, TN, for the human eyes, and Drs C. Dass and D. M. Desiderio of the Stout Neuroscience Laboratory, University of Tennessee,M emphis for their assistancei n the use of the mass spectrometer, which was obtained with the help of NIH Shared Equipment Grant RR0165 1. Supported by Public Health Service Grants EY 06540 (A.M.G) and EY 02665 (H.M.J.).
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1990/2
Y1 - 1990/2
N2 - Serine hydroxymethyltransferase (SHMT) is present in cultured rat, monkey and human lenses as shown by 15N-serine or 15N-glycine labeling studies. Following incubation with 15N-serine, the percent enrichment of 15N in glycine increases with time, and vice versa, demonstrating the presence of the enzyme, and the reversibility of the reaction in intact cultured lenses. Similar patterns of 15N enrichment were found in all three species, but lenses from young rats showed a higher percent enrichment than lenses obtained from older animals. Label from 15N-serine or 15N-glycine was also incorporated into a number of other amino acids, including aspartate, alanine, glutamate and proline. Conclusive evidence for the presence of SHMT in rat lens homogenates has been obtained by direct enzyme assay. The specific activity of rat lens SHMT was age dependent; approximately 2·4 units per mg protein in day old rats, declining to about 0·15 units per mg in adult animals. The higher specific activity observed in younger animals is consistent with the 15N labeling results obtained with cultured lenses. Lens SHMT has been partially characterized. In the presence of excess tetrahydrofolate the assay was essentially linear with increasing time. With serine as the substrate, the enzyme requires tetrahydrofolate for activity, the pH optimum is between pH 7·5 and 8·3, the Km for serine is about 0·25 mm, and the enzyme is inhibited by cycloserine. In conclusion, this study demonstrates the existence of SHMT in rat, monkey and human lenses. Rat lens specific activity has been shown to decrease with increasing age, and the enzyme has been partially characterized. Activity was detected by measuring the direct formation of the product, N5,N10-methylenetetrahydrofolate, and by following the reversible interconversion of 15N-labeled serine and glycine in intact cultured lenses.
AB - Serine hydroxymethyltransferase (SHMT) is present in cultured rat, monkey and human lenses as shown by 15N-serine or 15N-glycine labeling studies. Following incubation with 15N-serine, the percent enrichment of 15N in glycine increases with time, and vice versa, demonstrating the presence of the enzyme, and the reversibility of the reaction in intact cultured lenses. Similar patterns of 15N enrichment were found in all three species, but lenses from young rats showed a higher percent enrichment than lenses obtained from older animals. Label from 15N-serine or 15N-glycine was also incorporated into a number of other amino acids, including aspartate, alanine, glutamate and proline. Conclusive evidence for the presence of SHMT in rat lens homogenates has been obtained by direct enzyme assay. The specific activity of rat lens SHMT was age dependent; approximately 2·4 units per mg protein in day old rats, declining to about 0·15 units per mg in adult animals. The higher specific activity observed in younger animals is consistent with the 15N labeling results obtained with cultured lenses. Lens SHMT has been partially characterized. In the presence of excess tetrahydrofolate the assay was essentially linear with increasing time. With serine as the substrate, the enzyme requires tetrahydrofolate for activity, the pH optimum is between pH 7·5 and 8·3, the Km for serine is about 0·25 mm, and the enzyme is inhibited by cycloserine. In conclusion, this study demonstrates the existence of SHMT in rat, monkey and human lenses. Rat lens specific activity has been shown to decrease with increasing age, and the enzyme has been partially characterized. Activity was detected by measuring the direct formation of the product, N5,N10-methylenetetrahydrofolate, and by following the reversible interconversion of 15N-labeled serine and glycine in intact cultured lenses.
KW - glycine
KW - lens
KW - methionine synthesis
KW - serine
KW - serine hydroxymethyltransferase
KW - tetrahydrofolate
KW - transaminase
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U2 - 10.1016/0014-4835(90)90225-J
DO - 10.1016/0014-4835(90)90225-J
M3 - Article
C2 - 2311678
AN - SCOPUS:0025178659
SN - 0014-4835
VL - 50
SP - 149
EP - 155
JO - Experimental eye research
JF - Experimental eye research
IS - 2
ER -