TY - JOUR

T1 - Analysis of peptides and small proteins in preparations of horse milk exosomes, purified on anti-CD81-Sepharose

AU - Sedykh, Sergey E.

AU - Purvinish, Lada V.

AU - Burkova, Evgeniya E.

AU - Dmitrenok, Pavel S.

AU - Vlassov, Valentin V.

AU - Ryabchikova, Elena I.

AU - Nevinsky, Georgy A.

N1 - Funding Information: The research was supported by the Russian Science Foundation , project No. 18-74-10055 (to Sergey Sedykh); experiments in MALDI spectrometry were supported by the RFBR , project No. 18-04-00442_a (to Pavel Dmitrenok) and the Russian state-funded budget project of ICBFM SB RAS # АААА-А17-117020210023-1 (to Georgy Nevinsky). Publisher Copyright: © 2021 Elsevier Ltd Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/6

Y1 - 2021/6

N2 - Exosomes are nanovesicles (40–100 nm) involved in intracellular communication. Milk is a unique source of exosomes. In our previous papers, our group was the first to describe horse milk exosomes. Here we show how crude vesicle preparations were obtained from 18 horse milk samples by standard centrifugation, ultracentrifugation, and gel-filtration methods. Transmission electron microscopy and flow cytometry confirmed that exosomes contain tetraspanins on their surface. Affinity chromatography of vesicle preparations on anti-CD81-Sepharose produced extra-purified exosomes with more than 20 peptides and small proteins of 0.8–10 kDa as detected by MALDI-TOF mass spectrometry. The peptides are natural intrinsic components of horse milk exosomes and are not formed either as a result of or during sample preparation. We assume some of the hundreds and thousands of exosome proteins previously described by other authors in milk exosomes by highly sensitive liquid chromatography-mass spectrometry to have been misidentified as full-length proteins.

AB - Exosomes are nanovesicles (40–100 nm) involved in intracellular communication. Milk is a unique source of exosomes. In our previous papers, our group was the first to describe horse milk exosomes. Here we show how crude vesicle preparations were obtained from 18 horse milk samples by standard centrifugation, ultracentrifugation, and gel-filtration methods. Transmission electron microscopy and flow cytometry confirmed that exosomes contain tetraspanins on their surface. Affinity chromatography of vesicle preparations on anti-CD81-Sepharose produced extra-purified exosomes with more than 20 peptides and small proteins of 0.8–10 kDa as detected by MALDI-TOF mass spectrometry. The peptides are natural intrinsic components of horse milk exosomes and are not formed either as a result of or during sample preparation. We assume some of the hundreds and thousands of exosome proteins previously described by other authors in milk exosomes by highly sensitive liquid chromatography-mass spectrometry to have been misidentified as full-length proteins.

UR - http://www.scopus.com/inward/record.url?scp=85100623886&partnerID=8YFLogxK

U2 - 10.1016/j.idairyj.2021.104994

DO - 10.1016/j.idairyj.2021.104994

M3 - Article

AN - SCOPUS:85100623886

VL - 117

JO - International Dairy Journal

JF - International Dairy Journal

SN - 0958-6946

M1 - 104994

ER -