Lipoprotein lipase is active as a monomer

Anne P. Beigneux, Christopher M. Allan, Norma P. Sandoval, Geoffrey W. Cho, Patrick J. Heizer, Rachel S. Jung, Kimber Stanhope, Peter J Havel, Gabriel Birrane, Muthuraman Meiyappan, John E. Gill Iv, Masami Murakami, Kazuya Miyashita, Katsuyuki Nakajima, Michael Ploug, Loren G. Fong, Stephen G. Young

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Lipoprotein lipase (LPL), the enzyme that hydrolyzes triglycerides in plasma lipoproteins, is assumed to be active only as a homodimer. In support of this idea, several groups have reported that the size of LPL, as measured by density gradient ultracentrifugation, is ∼110 kDa, twice the size of LPL monomers (∼55 kDa). Of note, however, in those studies the LPL had been incubated with heparin, a polyanionic substance that binds and stabilizes LPL. Here we revisited the assumption that LPL is active only as a homodimer. When freshly secreted human LPL (or purified preparations of LPL) was subjected to density gradient ultracentrifugation (in the absence of heparin), LPL mass and activity peaks exhibited the size expected of monomers (near the 66-kDa albumin standard). GPIHBP1-bound LPL also exhibited the size expected for a monomer. In the presence of heparin, LPL size increased, overlapping with a 97.2-kDa standard. We also used density gradient ultracentrifugation to characterize the LPL within the high-salt and low-salt peaks from a heparin- Sepharose column. The catalytically active LPL within the high-salt peak exhibited the size of monomers, whereas most of the inactive LPL in the low-salt peak was at the bottom of the tube (in aggregates). Consistent with those findings, the LPL in the low-salt peak, but not that in the high-salt peak, was easily detectable with single mAb sandwich ELISAs, in which LPL is captured and detected with the same antibody. We conclude that catalytically active LPL can exist in a monomeric state.

Original languageEnglish (US)
Pages (from-to)6319-6328
Number of pages10
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number13
DOIs
StatePublished - Jan 1 2019

Fingerprint

Lipoprotein Lipase
Salts
Ultracentrifugation
Heparin

Keywords

  • Lipase
  • Lipolysis
  • Triglycerides

ASJC Scopus subject areas

  • General

Cite this

Beigneux, A. P., Allan, C. M., Sandoval, N. P., Cho, G. W., Heizer, P. J., Jung, R. S., ... Young, S. G. (2019). Lipoprotein lipase is active as a monomer. Proceedings of the National Academy of Sciences of the United States of America, 116(13), 6319-6328. https://doi.org/10.1073/pnas.1900983116

Lipoprotein lipase is active as a monomer. / Beigneux, Anne P.; Allan, Christopher M.; Sandoval, Norma P.; Cho, Geoffrey W.; Heizer, Patrick J.; Jung, Rachel S.; Stanhope, Kimber; Havel, Peter J; Birrane, Gabriel; Meiyappan, Muthuraman; Gill Iv, John E.; Murakami, Masami; Miyashita, Kazuya; Nakajima, Katsuyuki; Ploug, Michael; Fong, Loren G.; Young, Stephen G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 13, 01.01.2019, p. 6319-6328.

Research output: Contribution to journalArticle

Beigneux, AP, Allan, CM, Sandoval, NP, Cho, GW, Heizer, PJ, Jung, RS, Stanhope, K, Havel, PJ, Birrane, G, Meiyappan, M, Gill Iv, JE, Murakami, M, Miyashita, K, Nakajima, K, Ploug, M, Fong, LG & Young, SG 2019, 'Lipoprotein lipase is active as a monomer', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 13, pp. 6319-6328. https://doi.org/10.1073/pnas.1900983116
Beigneux, Anne P. ; Allan, Christopher M. ; Sandoval, Norma P. ; Cho, Geoffrey W. ; Heizer, Patrick J. ; Jung, Rachel S. ; Stanhope, Kimber ; Havel, Peter J ; Birrane, Gabriel ; Meiyappan, Muthuraman ; Gill Iv, John E. ; Murakami, Masami ; Miyashita, Kazuya ; Nakajima, Katsuyuki ; Ploug, Michael ; Fong, Loren G. ; Young, Stephen G. / Lipoprotein lipase is active as a monomer. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 13. pp. 6319-6328.
@article{3a1789760a6b4fd5b4595a7fb0695789,
title = "Lipoprotein lipase is active as a monomer",
abstract = "Lipoprotein lipase (LPL), the enzyme that hydrolyzes triglycerides in plasma lipoproteins, is assumed to be active only as a homodimer. In support of this idea, several groups have reported that the size of LPL, as measured by density gradient ultracentrifugation, is ∼110 kDa, twice the size of LPL monomers (∼55 kDa). Of note, however, in those studies the LPL had been incubated with heparin, a polyanionic substance that binds and stabilizes LPL. Here we revisited the assumption that LPL is active only as a homodimer. When freshly secreted human LPL (or purified preparations of LPL) was subjected to density gradient ultracentrifugation (in the absence of heparin), LPL mass and activity peaks exhibited the size expected of monomers (near the 66-kDa albumin standard). GPIHBP1-bound LPL also exhibited the size expected for a monomer. In the presence of heparin, LPL size increased, overlapping with a 97.2-kDa standard. We also used density gradient ultracentrifugation to characterize the LPL within the high-salt and low-salt peaks from a heparin- Sepharose column. The catalytically active LPL within the high-salt peak exhibited the size of monomers, whereas most of the inactive LPL in the low-salt peak was at the bottom of the tube (in aggregates). Consistent with those findings, the LPL in the low-salt peak, but not that in the high-salt peak, was easily detectable with single mAb sandwich ELISAs, in which LPL is captured and detected with the same antibody. We conclude that catalytically active LPL can exist in a monomeric state.",
keywords = "Lipase, Lipolysis, Triglycerides",
author = "Beigneux, {Anne P.} and Allan, {Christopher M.} and Sandoval, {Norma P.} and Cho, {Geoffrey W.} and Heizer, {Patrick J.} and Jung, {Rachel S.} and Kimber Stanhope and Havel, {Peter J} and Gabriel Birrane and Muthuraman Meiyappan and {Gill Iv}, {John E.} and Masami Murakami and Kazuya Miyashita and Katsuyuki Nakajima and Michael Ploug and Fong, {Loren G.} and Young, {Stephen G.}",
year = "2019",
month = "1",
day = "1",
doi = "10.1073/pnas.1900983116",
language = "English (US)",
volume = "116",
pages = "6319--6328",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "13",

}

TY - JOUR

T1 - Lipoprotein lipase is active as a monomer

AU - Beigneux, Anne P.

AU - Allan, Christopher M.

AU - Sandoval, Norma P.

AU - Cho, Geoffrey W.

AU - Heizer, Patrick J.

AU - Jung, Rachel S.

AU - Stanhope, Kimber

AU - Havel, Peter J

AU - Birrane, Gabriel

AU - Meiyappan, Muthuraman

AU - Gill Iv, John E.

AU - Murakami, Masami

AU - Miyashita, Kazuya

AU - Nakajima, Katsuyuki

AU - Ploug, Michael

AU - Fong, Loren G.

AU - Young, Stephen G.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Lipoprotein lipase (LPL), the enzyme that hydrolyzes triglycerides in plasma lipoproteins, is assumed to be active only as a homodimer. In support of this idea, several groups have reported that the size of LPL, as measured by density gradient ultracentrifugation, is ∼110 kDa, twice the size of LPL monomers (∼55 kDa). Of note, however, in those studies the LPL had been incubated with heparin, a polyanionic substance that binds and stabilizes LPL. Here we revisited the assumption that LPL is active only as a homodimer. When freshly secreted human LPL (or purified preparations of LPL) was subjected to density gradient ultracentrifugation (in the absence of heparin), LPL mass and activity peaks exhibited the size expected of monomers (near the 66-kDa albumin standard). GPIHBP1-bound LPL also exhibited the size expected for a monomer. In the presence of heparin, LPL size increased, overlapping with a 97.2-kDa standard. We also used density gradient ultracentrifugation to characterize the LPL within the high-salt and low-salt peaks from a heparin- Sepharose column. The catalytically active LPL within the high-salt peak exhibited the size of monomers, whereas most of the inactive LPL in the low-salt peak was at the bottom of the tube (in aggregates). Consistent with those findings, the LPL in the low-salt peak, but not that in the high-salt peak, was easily detectable with single mAb sandwich ELISAs, in which LPL is captured and detected with the same antibody. We conclude that catalytically active LPL can exist in a monomeric state.

AB - Lipoprotein lipase (LPL), the enzyme that hydrolyzes triglycerides in plasma lipoproteins, is assumed to be active only as a homodimer. In support of this idea, several groups have reported that the size of LPL, as measured by density gradient ultracentrifugation, is ∼110 kDa, twice the size of LPL monomers (∼55 kDa). Of note, however, in those studies the LPL had been incubated with heparin, a polyanionic substance that binds and stabilizes LPL. Here we revisited the assumption that LPL is active only as a homodimer. When freshly secreted human LPL (or purified preparations of LPL) was subjected to density gradient ultracentrifugation (in the absence of heparin), LPL mass and activity peaks exhibited the size expected of monomers (near the 66-kDa albumin standard). GPIHBP1-bound LPL also exhibited the size expected for a monomer. In the presence of heparin, LPL size increased, overlapping with a 97.2-kDa standard. We also used density gradient ultracentrifugation to characterize the LPL within the high-salt and low-salt peaks from a heparin- Sepharose column. The catalytically active LPL within the high-salt peak exhibited the size of monomers, whereas most of the inactive LPL in the low-salt peak was at the bottom of the tube (in aggregates). Consistent with those findings, the LPL in the low-salt peak, but not that in the high-salt peak, was easily detectable with single mAb sandwich ELISAs, in which LPL is captured and detected with the same antibody. We conclude that catalytically active LPL can exist in a monomeric state.

KW - Lipase

KW - Lipolysis

KW - Triglycerides

UR - http://www.scopus.com/inward/record.url?scp=85063936170&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85063936170&partnerID=8YFLogxK

U2 - 10.1073/pnas.1900983116

DO - 10.1073/pnas.1900983116

M3 - Article

C2 - 30850549

AN - SCOPUS:85063936170

VL - 116

SP - 6319

EP - 6328

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 13

ER -