Identification of a novel alternatively spliced isoform of antithrombin containing an additional RCL-like loop

Sayeed ur Rehman, Shazia Ashraf, Shahzaib Ahamad, Tarique Sarwar, Mohamad Amir Husain, Parvez Ahmad, Mohammad Tabish, Mohamad Aman Jairajpuri

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Antithrombin (AT3) is one of the most important inhibitors of blood coagulation proteases that belong to the serpin family of protease inhibitors. In this study, a novel alternatively spliced isoform of AT3 was identified, both at transcript and protein level. This novel transcript contains an additional region in the continuation of exon 3b that was included in the transcript due to use of an alternate 5’ splice site. The existence of the novel transcript was confirmed in human brain and liver through RT-PCR. An analysis of the complete transcript indicated that the native reactive centre loop (RCL) of AT3 is maintained; however the novel amino acid sequence projects out as an additional loop as evident from MD simulation studies. A unique amino acid sequence present in the novel isoform was used for the development of polyclonal antibody. The expression of novel isoform was confirmed in human brain and liver tissue using Western blot analysis. Interestingly an alignment of RCL like domain with other inhibitory serpins showed significant similarity with the neuroserpin RCL. To the best of our knowledge, this is the first evidence of alternatively spliced AT3 sequence containing an additional loop and could have physiological relevance.

Original languageEnglish (US)
Pages (from-to)421-426
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume517
Issue number3
DOIs
StatePublished - Sep 24 2019
Externally publishedYes

Keywords

  • Alternative splicing
  • Antithrombin
  • Neuroserpin
  • Novel isoform
  • Reactive centre loop

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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