Combining different proteomic approaches to resolve complexity of the milk protein fraction of dromedary, Bactrian camels and hybrids, from different regions of Kazakhstan

PLoS One. 2018 May 10;13(5):e0197026. doi: 10.1371/journal.pone.0197026. eCollection 2018.

Abstract

Nutritional suitability of milk is not only related to gross composition, but is also strongly affected by the microheterogeniety of the protein fraction. Hence, to go further into the evaluation of the potential suitability of non-bovine milks in human/infant nutrition it is necessary to have a detailed characterization of their protein components. Combining proven proteomic approaches (SDS-PAGE, LC-MS/MS and LC-ESI-MS) and cDNA sequencing, we provide here in depth characterization of the milk protein fraction of dromedary and Bactrian camels, and their hybrids, from different regions of Kazakhstan. A total 391 functional groups of proteins were identified from 8 camel milk samples. A detailed characterization of 50 protein molecules, relating to genetic variants and isoforms arising from post-translational modifications and alternative splicing events, belonging to nine protein families (κ-, αs1-, αs2-, β-; and γ-CN, WAP, α-LAC, PGRP, CSA/LPO) was achieved by LC-ESI-MS. The presence of two unknown proteins UP1 (22,939 Da) and UP2 (23,046 Da) was also reported as well as the existence of a β-CN short isoform (946 Da lighter than the full-length β-CN), arising very likely in both genetic variants (A and B) from proteolysis by plasmin. In addition, we report, for the first time to our knowledge, the occurrence of a αs2-CN phosphorylation isoform with 12P groups within two recognition motifs, suggesting thereby the existence of two kinase systems involved in the phosphorylation of caseins in the mammary gland. Finally, we demonstrate that genetic variants, which hitherto seemed to be species- specific (e.g. β-CN A for Bactrian and β-CN B for dromedary), are in fact present both in Camel dromedarius and C. bactrianus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Camelus / genetics
  • Camelus / metabolism*
  • Chimera / genetics
  • Chimera / metabolism*
  • Female
  • Kazakhstan
  • Mass Spectrometry
  • Milk Proteins / metabolism*
  • Proteomics*

Substances

  • Milk Proteins

Grants and funding

The study was carried out within the Bolashak International Scholarship of the first author, funded by the JSC «Center for International Programs» (Kazakhstan). The research was partly supported by a grant from the Ministry of Education and Science of the Republic of Kazakhstan under name “Proteomic investigation of camel milk” #1729/GF4, which is duly appreciated. INRA (Jouy-en-Josas, France) for providing necessary facilities and technical support.