A method for assaying the activity of the endopeptidase which excises the nonhelical carboxyterminal extensions from type I procollagen

Efrat Kessler; Burton Goldberg

doi:10.1016/0003-2697(78)90770-4

A method for assaying the activity of the endopeptidase which excises the nonhelical carboxyterminal extensions from type I procollagen

Efrat Kessler^*, Burton Goldberg

^*Corresponding author for this work

New York University

Research output: Contribution to journal › Article › peer-review

28 Scopus citations

Abstract

A new method for measuring the rate of enzymatic excision of the carboxy-terminal, nonhelical fragment from type I procollagen is presented. Human procollagen containing [³H]tryptophan-labeled carboxy-terminal extensions was used as the substrate, and the enzyme was derived from the culture medium of mouse 3T6 fibroblasts. Incubation mixtures of substrate with enzyme were made 25% in ethanol which left the excised radiolabeled carboxy-terminal fragment in solution, whereas all other radiolabeled components were precipitated. Enzymatic activity was measured by radioactive counting of the ethanol supernatant. The assay is simple, rapid, sensitive and generates valid kinetic data.

Original language	English
Pages (from-to)	463-469
Number of pages	7
Journal	Analytical Biochemistry
Volume	86
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(78)90770-4
State	Published - 1 Jun 1978
Externally published	Yes

Funding

Funders	Funder number
National Institutes of Health	5 ROI HLl7551, 5 SO7 RR05399

Access to Document

10.1016/0003-2697(78)90770-4

Cite this

@article{4862341ae63143a0b665e2abd92a147f,

title = "A method for assaying the activity of the endopeptidase which excises the nonhelical carboxyterminal extensions from type I procollagen",

abstract = "A new method for measuring the rate of enzymatic excision of the carboxy-terminal, nonhelical fragment from type I procollagen is presented. Human procollagen containing [3H]tryptophan-labeled carboxy-terminal extensions was used as the substrate, and the enzyme was derived from the culture medium of mouse 3T6 fibroblasts. Incubation mixtures of substrate with enzyme were made 25% in ethanol which left the excised radiolabeled carboxy-terminal fragment in solution, whereas all other radiolabeled components were precipitated. Enzymatic activity was measured by radioactive counting of the ethanol supernatant. The assay is simple, rapid, sensitive and generates valid kinetic data.",

author = "Efrat Kessler and Burton Goldberg",

note = "Funding Information: This work was supported by NIH Grants 5 ROI HLl7551 and 5 SO7 RR05399. The authors gratefully acknowledge the excellent assistance of Sheila Heitner.",

year = "1978",

month = jun,

day = "1",

doi = "10.1016/0003-2697(78)90770-4",

language = "אנגלית",

volume = "86",

pages = "463--469",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

number = "2",

}

TY - JOUR

T1 - A method for assaying the activity of the endopeptidase which excises the nonhelical carboxyterminal extensions from type I procollagen

AU - Kessler, Efrat

AU - Goldberg, Burton

N1 - Funding Information: This work was supported by NIH Grants 5 ROI HLl7551 and 5 SO7 RR05399. The authors gratefully acknowledge the excellent assistance of Sheila Heitner.

PY - 1978/6/1

Y1 - 1978/6/1

N2 - A new method for measuring the rate of enzymatic excision of the carboxy-terminal, nonhelical fragment from type I procollagen is presented. Human procollagen containing [3H]tryptophan-labeled carboxy-terminal extensions was used as the substrate, and the enzyme was derived from the culture medium of mouse 3T6 fibroblasts. Incubation mixtures of substrate with enzyme were made 25% in ethanol which left the excised radiolabeled carboxy-terminal fragment in solution, whereas all other radiolabeled components were precipitated. Enzymatic activity was measured by radioactive counting of the ethanol supernatant. The assay is simple, rapid, sensitive and generates valid kinetic data.

AB - A new method for measuring the rate of enzymatic excision of the carboxy-terminal, nonhelical fragment from type I procollagen is presented. Human procollagen containing [3H]tryptophan-labeled carboxy-terminal extensions was used as the substrate, and the enzyme was derived from the culture medium of mouse 3T6 fibroblasts. Incubation mixtures of substrate with enzyme were made 25% in ethanol which left the excised radiolabeled carboxy-terminal fragment in solution, whereas all other radiolabeled components were precipitated. Enzymatic activity was measured by radioactive counting of the ethanol supernatant. The assay is simple, rapid, sensitive and generates valid kinetic data.

UR - http://www.scopus.com/inward/record.url?scp=0018169011&partnerID=8YFLogxK

U2 - 10.1016/0003-2697(78)90770-4

DO - 10.1016/0003-2697(78)90770-4

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

AN - SCOPUS:0018169011

SN - 0003-2697

VL - 86

SP - 463

EP - 469

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

A method for assaying the activity of the endopeptidase which excises the nonhelical carboxyterminal extensions from type I procollagen

Abstract

Funding

Access to Document

Other files and links

Fingerprint

Cite this