Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)
Fibronectin type III domain containing 5 (FNDC5) is a transmembrane protein. Upon cleavage, it yields a peptide called irisin that is supposedly bind to an unknown receptor and facilitates browning of white adipose tissue (WAT). Increased levels of irisin are associated with increased levels of ener...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-12-01
|
| Series: | Journal of Genetic Engineering and Biotechnology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1687157X18300672 |
| _version_ | 1797203429540495360 |
|---|---|
| author | Kalpana Panati Venkata Ramireddy Narala Vydyanath R. Narasimha Madhavi Derangula Venkat R.R. Arva Tatireddigari Suneetha Yeguvapalli |
| author_facet | Kalpana Panati Venkata Ramireddy Narala Vydyanath R. Narasimha Madhavi Derangula Venkat R.R. Arva Tatireddigari Suneetha Yeguvapalli |
| author_sort | Kalpana Panati |
| collection | DOAJ |
| description | Fibronectin type III domain containing 5 (FNDC5) is a transmembrane protein. Upon cleavage, it yields a peptide called irisin that is supposedly bind to an unknown receptor and facilitates browning of white adipose tissue (WAT). Increased levels of irisin are associated with increased levels of energy expenditure markers PGC-1α, UCP-1, besides abundance of beige adipocytes in WAT. Though varied sizes of irisin were reported in humans and rodents it is not yet clear about the actual size of the irisin produced physiologically. Hence, we cloned and expressed human irisin (32–143 aa of FNDC5) in Escherichia coli based on the proposed cleavage site that yields 12.5 kDa peptide to study its antigenicity and other biological functions in vitro. We purified recombinant human irisin (rh-irisin) to 95% homogeneity with simple purification method with a yield of 25 mg/g wet cell pellet. rh-irisin has been detected by commercially available antibodies from different sources with similar antigenicity. Biological activity of the rh-irisin was confirmed by using 3T3-L1 pre-adipocyte differentiation by Oil red O staining. Further, rh-irisin treatment on pre-adipocytes showed increased expression of markers associated with energy expenditure. As it is involved in energy expenditure process, it could be considered as potential therapeutic option for various metabolic diseases. Keywords: Beige adipose tissue, Energy expenditure, FNDC5, Irisin, Obesity, UCP-1 |
| first_indexed | 2024-04-24T08:19:12Z |
| format | Article |
| id | doaj.art-5d0b31275f51446098be4a8e2ca2a951 |
| institution | Directory Open Access Journal |
| issn | 1687-157X |
| language | English |
| last_indexed | 2024-04-24T08:19:12Z |
| publishDate | 2018-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Genetic Engineering and Biotechnology |
| spelling | doaj.art-5d0b31275f51446098be4a8e2ca2a9512024-04-17T02:14:11ZengElsevierJournal of Genetic Engineering and Biotechnology1687-157X2018-12-01162459466Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)Kalpana Panati0Venkata Ramireddy Narala1Vydyanath R. Narasimha2Madhavi Derangula3Venkat R.R. Arva Tatireddigari4Suneetha Yeguvapalli5Department of Biotechnology, Government College for Men, Kadapa, AP 516 004, India; Department of Biotechnology, Sri Venkateswara University, Tirupati, AP 517 502, India; Corresponding authors at: Department of Biotechnology, Government College for Men, Kadapa 516 004, AP, India (K. Panati).Department of Zoology, Yogi Vemana University, Kadapa, AP 516 005, IndiaDepartment of Zoology, Yogi Vemana University, Kadapa, AP 516 005, IndiaDepartment of Zoology, Yogi Vemana University, Kadapa, AP 516 005, IndiaDepartment of Zoology, Yogi Vemana University, Kadapa, AP 516 005, IndiaDepartment of Zoology, Sri Venkateswara University, Tirupati, AP 517 502, India; Corresponding authors at: Department of Biotechnology, Government College for Men, Kadapa 516 004, AP, India (K. Panati).Fibronectin type III domain containing 5 (FNDC5) is a transmembrane protein. Upon cleavage, it yields a peptide called irisin that is supposedly bind to an unknown receptor and facilitates browning of white adipose tissue (WAT). Increased levels of irisin are associated with increased levels of energy expenditure markers PGC-1α, UCP-1, besides abundance of beige adipocytes in WAT. Though varied sizes of irisin were reported in humans and rodents it is not yet clear about the actual size of the irisin produced physiologically. Hence, we cloned and expressed human irisin (32–143 aa of FNDC5) in Escherichia coli based on the proposed cleavage site that yields 12.5 kDa peptide to study its antigenicity and other biological functions in vitro. We purified recombinant human irisin (rh-irisin) to 95% homogeneity with simple purification method with a yield of 25 mg/g wet cell pellet. rh-irisin has been detected by commercially available antibodies from different sources with similar antigenicity. Biological activity of the rh-irisin was confirmed by using 3T3-L1 pre-adipocyte differentiation by Oil red O staining. Further, rh-irisin treatment on pre-adipocytes showed increased expression of markers associated with energy expenditure. As it is involved in energy expenditure process, it could be considered as potential therapeutic option for various metabolic diseases. Keywords: Beige adipose tissue, Energy expenditure, FNDC5, Irisin, Obesity, UCP-1http://www.sciencedirect.com/science/article/pii/S1687157X18300672 |
| spellingShingle | Kalpana Panati Venkata Ramireddy Narala Vydyanath R. Narasimha Madhavi Derangula Venkat R.R. Arva Tatireddigari Suneetha Yeguvapalli Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa) Journal of Genetic Engineering and Biotechnology |
| title | Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa) |
| title_full | Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa) |
| title_fullStr | Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa) |
| title_full_unstemmed | Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa) |
| title_short | Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa) |
| title_sort | expression purification and biological characterisation of recombinant human irisin 12 5 kda |
| url | http://www.sciencedirect.com/science/article/pii/S1687157X18300672 |
| work_keys_str_mv | AT kalpanapanati expressionpurificationandbiologicalcharacterisationofrecombinanthumanirisin125kda AT venkataramireddynarala expressionpurificationandbiologicalcharacterisationofrecombinanthumanirisin125kda AT vydyanathrnarasimha expressionpurificationandbiologicalcharacterisationofrecombinanthumanirisin125kda AT madhaviderangula expressionpurificationandbiologicalcharacterisationofrecombinanthumanirisin125kda AT venkatrrarvatatireddigari expressionpurificationandbiologicalcharacterisationofrecombinanthumanirisin125kda AT suneethayeguvapalli expressionpurificationandbiologicalcharacterisationofrecombinanthumanirisin125kda |