Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk

Jagadeesh Janjanam; Manu Jamwal; Surender V. Singh; Saravanan Kumar; Aswini Kumar Panigrahi; Gururao Hariprasad; Manoj Kumar Jena; Vijay R. Anand; Sudarshan Suresh Kumar; Jai Kumar Kaushik; Ajaykumar Dang; Manishi Mukesh; Bishnu Prasad Mishra; Alagiri Srinivasan; Vanga Siva Belum Reddy; Ashok Kumar Mohanty

doi:10.1002/pmic.201300031

Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk

Jagadeesh Janjanam, Manu Jamwal, Surender V. Singh, Saravanan Kumar, Aswini Kumar Panigrahi, Gururao Hariprasad, Manoj Kumar Jena, Vijay R. Anand, Sudarshan Suresh Kumar, Jai Kumar Kaushik, Ajaykumar Dang, Manishi Mukesh, Bishnu Prasad Mishra, Alagiri Srinivasan, Vanga Siva Belum Reddy, Ashok Kumar Mohanty

Bioscience Core Laboratory

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

26 Scopus citations

Abstract

Mammary gland is made up of a branching network of ducts that end in alveoli. Terminally differentiated mammary epithelial cells (MECs) constitute the innermost layer of aveoli. They are milk-secreting cuboidal cells that secrete milk proteins during lactation. Little is known about the expression profile of proteins in the metabolically active MECs during lactation or their functional role in the lactation process. In the present investigation, we have reported the proteome map of MECs in lactating cows using 2DE MALDI-TOF/TOF MS and 1D-Gel-LC-MS/MS. MECs were isolated from milk using immunomagnetic beads and confirmed by RT-PCR and Western blotting. The 1D-Gel-LC-MS/MS and 2DE-MS/MS based approaches led to identification of 431 and 134 proteins, respectively, with a total of 497 unique proteins. Proteins identified in this study were clustered into functional groups using bioinformatics tools. Pathway analysis of the identified proteins revealed 28 pathways (p < 0.05) providing evidence for involvement of various proteins in lactation function. This study further provides experimental evidence for the presence of many proteins that have been predicted in annotated bovine genome. The data generated further provide a set of bovine MEC-specific proteins that will help the researchers to understand the molecular events taking place during lactation. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Original language	English (US)
Pages (from-to)	3189-3204
Number of pages	16
Journal	PROTEOMICS
Volume	13
Issue number	21
DOIs	https://doi.org/10.1002/pmic.201300031
State	Published - Oct 1 2013

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: The research was supported by National Agriculture Innovation Project (component 4), Indian Council of Agricultural Research (ICAR), Government of India. A part of the MS data was collected at Seattle Biomed, Seattle, USA. J.J. received a junior research fellowship from the council of scientific research (CSIR), Government of India. We acknowledge the use of computational facility BTIS, sub center, NDRI.

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1002/pmic.201300031

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Janjanam, J., Jamwal, M., Singh, S. V., Kumar, S., Panigrahi, A. K., Hariprasad, G., Jena, M. K., Anand, V. R., Kumar, S. S., Kaushik, J. K., Dang, A., Mukesh, M., Mishra, B. P., Srinivasan, A., Reddy, V. S. B., & Mohanty, A. K. (2013). Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk. PROTEOMICS, 13(21), 3189-3204. https://doi.org/10.1002/pmic.201300031

@article{1a99759c2d3f475eb4a5f92bb7f62dd4,

title = "Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk",

abstract = "Mammary gland is made up of a branching network of ducts that end in alveoli. Terminally differentiated mammary epithelial cells (MECs) constitute the innermost layer of aveoli. They are milk-secreting cuboidal cells that secrete milk proteins during lactation. Little is known about the expression profile of proteins in the metabolically active MECs during lactation or their functional role in the lactation process. In the present investigation, we have reported the proteome map of MECs in lactating cows using 2DE MALDI-TOF/TOF MS and 1D-Gel-LC-MS/MS. MECs were isolated from milk using immunomagnetic beads and confirmed by RT-PCR and Western blotting. The 1D-Gel-LC-MS/MS and 2DE-MS/MS based approaches led to identification of 431 and 134 proteins, respectively, with a total of 497 unique proteins. Proteins identified in this study were clustered into functional groups using bioinformatics tools. Pathway analysis of the identified proteins revealed 28 pathways (p < 0.05) providing evidence for involvement of various proteins in lactation function. This study further provides experimental evidence for the presence of many proteins that have been predicted in annotated bovine genome. The data generated further provide a set of bovine MEC-specific proteins that will help the researchers to understand the molecular events taking place during lactation. {\textcopyright} 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.",

author = "Jagadeesh Janjanam and Manu Jamwal and Singh, {Surender V.} and Saravanan Kumar and Panigrahi, {Aswini Kumar} and Gururao Hariprasad and Jena, {Manoj Kumar} and Anand, {Vijay R.} and Kumar, {Sudarshan Suresh} and Kaushik, {Jai Kumar} and Ajaykumar Dang and Manishi Mukesh and Mishra, {Bishnu Prasad} and Alagiri Srinivasan and Reddy, {Vanga Siva Belum} and Mohanty, {Ashok Kumar}",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: The research was supported by National Agriculture Innovation Project (component 4), Indian Council of Agricultural Research (ICAR), Government of India. A part of the MS data was collected at Seattle Biomed, Seattle, USA. J.J. received a junior research fellowship from the council of scientific research (CSIR), Government of India. We acknowledge the use of computational facility BTIS, sub center, NDRI.",

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month = oct,

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doi = "10.1002/pmic.201300031",

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Janjanam, J, Jamwal, M, Singh, SV, Kumar, S, Panigrahi, AK, Hariprasad, G, Jena, MK, Anand, VR, Kumar, SS, Kaushik, JK, Dang, A, Mukesh, M, Mishra, BP, Srinivasan, A, Reddy, VSB & Mohanty, AK 2013, 'Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk', PROTEOMICS, vol. 13, no. 21, pp. 3189-3204. https://doi.org/10.1002/pmic.201300031

TY - JOUR

T1 - Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk

AU - Janjanam, Jagadeesh

AU - Jamwal, Manu

AU - Singh, Surender V.

AU - Kumar, Saravanan

AU - Panigrahi, Aswini Kumar

AU - Hariprasad, Gururao

AU - Jena, Manoj Kumar

AU - Anand, Vijay R.

AU - Kumar, Sudarshan Suresh

AU - Kaushik, Jai Kumar

AU - Dang, Ajaykumar

AU - Mukesh, Manishi

AU - Mishra, Bishnu Prasad

AU - Srinivasan, Alagiri

AU - Reddy, Vanga Siva Belum

AU - Mohanty, Ashok Kumar

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: The research was supported by National Agriculture Innovation Project (component 4), Indian Council of Agricultural Research (ICAR), Government of India. A part of the MS data was collected at Seattle Biomed, Seattle, USA. J.J. received a junior research fellowship from the council of scientific research (CSIR), Government of India. We acknowledge the use of computational facility BTIS, sub center, NDRI.

PY - 2013/10/1

Y1 - 2013/10/1

N2 - Mammary gland is made up of a branching network of ducts that end in alveoli. Terminally differentiated mammary epithelial cells (MECs) constitute the innermost layer of aveoli. They are milk-secreting cuboidal cells that secrete milk proteins during lactation. Little is known about the expression profile of proteins in the metabolically active MECs during lactation or their functional role in the lactation process. In the present investigation, we have reported the proteome map of MECs in lactating cows using 2DE MALDI-TOF/TOF MS and 1D-Gel-LC-MS/MS. MECs were isolated from milk using immunomagnetic beads and confirmed by RT-PCR and Western blotting. The 1D-Gel-LC-MS/MS and 2DE-MS/MS based approaches led to identification of 431 and 134 proteins, respectively, with a total of 497 unique proteins. Proteins identified in this study were clustered into functional groups using bioinformatics tools. Pathway analysis of the identified proteins revealed 28 pathways (p < 0.05) providing evidence for involvement of various proteins in lactation function. This study further provides experimental evidence for the presence of many proteins that have been predicted in annotated bovine genome. The data generated further provide a set of bovine MEC-specific proteins that will help the researchers to understand the molecular events taking place during lactation. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

AB - Mammary gland is made up of a branching network of ducts that end in alveoli. Terminally differentiated mammary epithelial cells (MECs) constitute the innermost layer of aveoli. They are milk-secreting cuboidal cells that secrete milk proteins during lactation. Little is known about the expression profile of proteins in the metabolically active MECs during lactation or their functional role in the lactation process. In the present investigation, we have reported the proteome map of MECs in lactating cows using 2DE MALDI-TOF/TOF MS and 1D-Gel-LC-MS/MS. MECs were isolated from milk using immunomagnetic beads and confirmed by RT-PCR and Western blotting. The 1D-Gel-LC-MS/MS and 2DE-MS/MS based approaches led to identification of 431 and 134 proteins, respectively, with a total of 497 unique proteins. Proteins identified in this study were clustered into functional groups using bioinformatics tools. Pathway analysis of the identified proteins revealed 28 pathways (p < 0.05) providing evidence for involvement of various proteins in lactation function. This study further provides experimental evidence for the presence of many proteins that have been predicted in annotated bovine genome. The data generated further provide a set of bovine MEC-specific proteins that will help the researchers to understand the molecular events taking place during lactation. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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UR - http://doi.wiley.com/10.1002/pmic.201300031

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U2 - 10.1002/pmic.201300031

DO - 10.1002/pmic.201300031

M3 - Article

C2 - 24030930

SN - 1615-9853

VL - 13

SP - 3189

EP - 3204

JO - PROTEOMICS

JF - PROTEOMICS

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ER -

Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk

Abstract

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ASJC Scopus subject areas

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