Rapid enzyme assays investigating the variation in the glycolytic pathway in field-caught populations of Fundulus heteroclitus

Valerie A. Pierce, Douglas L Crawford

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Variation in enzyme expression may be important in evolutionary adaptation, yet is seldom studied. Furthermore, no studies have examined the expression of all enzymes in a defined metabolic pathway. Enzyme concentration is a measure of enzyme expression and was ascertained by assaying maximal activity. Presented here is an analysis of variation of maximal enzyme activity for all the enzymes in a single metabolic pathway, glycolysis, from three clinically distributed populations of the fish, Fundulus heteroclitus. Techniques for rapidly analyzing maximal enzyme activity for all the enzymes of an entire metabolic pathway from many individuals are described. The high degree of repeatability (mean coefficient of variation for replicates, 4.4%) and sensitivity (less than 3 mg of tissue is required to measure all 10 enzymes) of these assays demonstrate the utility of such an approach for analyzing variation among populations for a large numbers of enzymes. Results from these studies indicate that (1) the average coefficient of variation for all enzyme determinations within a population is 45.3% and (2) between populations, the activity of 5 of the 10 glycolytic enzymes are significantly different. This considerable variation occurs even in populations where there is little allelic variation. These data demonstrating substantial variation in enzyme expression support the idea that changes in gene regulation may be as important as, or even more important than, changes in biochemical kinetic parameters in evolutionary processes.

Original languageEnglish
Pages (from-to)315-330
Number of pages16
JournalBiochemical Genetics
Volume32
Issue number9-10
DOIs
StatePublished - Oct 1 1994
Externally publishedYes

Fingerprint

Fundulidae
Fundulus heteroclitus
Enzyme Assays
glycolysis
Assays
assay
enzyme
assays
Enzymes
enzymes
Population
Metabolic Networks and Pathways
biochemical pathways
Enzyme activity
enzyme activity
evolutionary adaptation
Kinetic parameters
Glycolysis
Gene expression
repeatability

Keywords

  • enzyme expression
  • Fundulus heteroclitus
  • glycolysis
  • maximal enzyme activity
  • microplate reader

ASJC Scopus subject areas

  • Genetics
  • Biochemistry

Cite this

Rapid enzyme assays investigating the variation in the glycolytic pathway in field-caught populations of Fundulus heteroclitus. / Pierce, Valerie A.; Crawford, Douglas L.

In: Biochemical Genetics, Vol. 32, No. 9-10, 01.10.1994, p. 315-330.

Research output: Contribution to journalArticle

@article{65c2f4a009b147fdb87e4f408ee3b5f1,
title = "Rapid enzyme assays investigating the variation in the glycolytic pathway in field-caught populations of Fundulus heteroclitus",
abstract = "Variation in enzyme expression may be important in evolutionary adaptation, yet is seldom studied. Furthermore, no studies have examined the expression of all enzymes in a defined metabolic pathway. Enzyme concentration is a measure of enzyme expression and was ascertained by assaying maximal activity. Presented here is an analysis of variation of maximal enzyme activity for all the enzymes in a single metabolic pathway, glycolysis, from three clinically distributed populations of the fish, Fundulus heteroclitus. Techniques for rapidly analyzing maximal enzyme activity for all the enzymes of an entire metabolic pathway from many individuals are described. The high degree of repeatability (mean coefficient of variation for replicates, 4.4{\%}) and sensitivity (less than 3 mg of tissue is required to measure all 10 enzymes) of these assays demonstrate the utility of such an approach for analyzing variation among populations for a large numbers of enzymes. Results from these studies indicate that (1) the average coefficient of variation for all enzyme determinations within a population is 45.3{\%} and (2) between populations, the activity of 5 of the 10 glycolytic enzymes are significantly different. This considerable variation occurs even in populations where there is little allelic variation. These data demonstrating substantial variation in enzyme expression support the idea that changes in gene regulation may be as important as, or even more important than, changes in biochemical kinetic parameters in evolutionary processes.",
keywords = "enzyme expression, Fundulus heteroclitus, glycolysis, maximal enzyme activity, microplate reader",
author = "Pierce, {Valerie A.} and Crawford, {Douglas L}",
year = "1994",
month = "10",
day = "1",
doi = "10.1007/BF02426894",
language = "English",
volume = "32",
pages = "315--330",
journal = "Biochemical Genetics",
issn = "0006-2928",
publisher = "Springer New York",
number = "9-10",

}

TY - JOUR

T1 - Rapid enzyme assays investigating the variation in the glycolytic pathway in field-caught populations of Fundulus heteroclitus

AU - Pierce, Valerie A.

AU - Crawford, Douglas L

PY - 1994/10/1

Y1 - 1994/10/1

N2 - Variation in enzyme expression may be important in evolutionary adaptation, yet is seldom studied. Furthermore, no studies have examined the expression of all enzymes in a defined metabolic pathway. Enzyme concentration is a measure of enzyme expression and was ascertained by assaying maximal activity. Presented here is an analysis of variation of maximal enzyme activity for all the enzymes in a single metabolic pathway, glycolysis, from three clinically distributed populations of the fish, Fundulus heteroclitus. Techniques for rapidly analyzing maximal enzyme activity for all the enzymes of an entire metabolic pathway from many individuals are described. The high degree of repeatability (mean coefficient of variation for replicates, 4.4%) and sensitivity (less than 3 mg of tissue is required to measure all 10 enzymes) of these assays demonstrate the utility of such an approach for analyzing variation among populations for a large numbers of enzymes. Results from these studies indicate that (1) the average coefficient of variation for all enzyme determinations within a population is 45.3% and (2) between populations, the activity of 5 of the 10 glycolytic enzymes are significantly different. This considerable variation occurs even in populations where there is little allelic variation. These data demonstrating substantial variation in enzyme expression support the idea that changes in gene regulation may be as important as, or even more important than, changes in biochemical kinetic parameters in evolutionary processes.

AB - Variation in enzyme expression may be important in evolutionary adaptation, yet is seldom studied. Furthermore, no studies have examined the expression of all enzymes in a defined metabolic pathway. Enzyme concentration is a measure of enzyme expression and was ascertained by assaying maximal activity. Presented here is an analysis of variation of maximal enzyme activity for all the enzymes in a single metabolic pathway, glycolysis, from three clinically distributed populations of the fish, Fundulus heteroclitus. Techniques for rapidly analyzing maximal enzyme activity for all the enzymes of an entire metabolic pathway from many individuals are described. The high degree of repeatability (mean coefficient of variation for replicates, 4.4%) and sensitivity (less than 3 mg of tissue is required to measure all 10 enzymes) of these assays demonstrate the utility of such an approach for analyzing variation among populations for a large numbers of enzymes. Results from these studies indicate that (1) the average coefficient of variation for all enzyme determinations within a population is 45.3% and (2) between populations, the activity of 5 of the 10 glycolytic enzymes are significantly different. This considerable variation occurs even in populations where there is little allelic variation. These data demonstrating substantial variation in enzyme expression support the idea that changes in gene regulation may be as important as, or even more important than, changes in biochemical kinetic parameters in evolutionary processes.

KW - enzyme expression

KW - Fundulus heteroclitus

KW - glycolysis

KW - maximal enzyme activity

KW - microplate reader

UR - http://www.scopus.com/inward/record.url?scp=0028608097&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028608097&partnerID=8YFLogxK

U2 - 10.1007/BF02426894

DO - 10.1007/BF02426894

M3 - Article

VL - 32

SP - 315

EP - 330

JO - Biochemical Genetics

JF - Biochemical Genetics

SN - 0006-2928

IS - 9-10

ER -