Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes

KARACİCEK, Bilge; Ceylan, Çağatay

Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes

Bilge KARACİCEK, (İzmir Yüksek Teknoloji Enstitüsü, Biyoteknoloji Programı, Urla, İzmir, Türkiye)

Çağatay CEYLAN (İzmir Yüksek Teknoloji Enstitüsü, Biyoteknoloji Programı, Urla, İzmir, Türkiye)

Türk Biyokimya Dergisi

2 0

Yıl: 2015 Cilt: 40 Sayı: 1 Sayfa Aralığı: 81 - 87 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes

Öz:

Bu çalışmada alkol-su karışımlarında pH tarafından tetiklenen sığır pankreatik tripsin jelleşmesi ve agregasyonu faz değişiklikleri üzerine üç değişik fizikokimyasal değişkenin yapısal ve fonksiyonel etkileri çalışılmıştır.Metod: Tripsin jelleşmesi ters tüp metodu ile çalışılmıştır. Tripsin stabilitesi tripsin enzim aktivitesi tayiniyle çalışılmıştır. Protein ikinci yapı değişiklikleri FTIR spektroskopisi metodu ile gözlenmiştir. Jel ve agregat makroyapıları ve morfolojileri taramalı elektron mikroskopisi yöntemiyle incelenmiştir.Bulgular: NaOH ve CaCl2 eksikliğinde çözelti fazı gözlenmiştir. Jel fazı bu ikisinden birinin eksikliğinde gözlenmiştir. Agregat fazı ise bu iki ajanın birlikte varlığında tripsin konantrasyonuna bağlı olarak gözlenmiştir. Tripsin dayanıklılık çalışmaları jelleşmede yaklaşık olarak 53% ve agregasyonda ise 32% özgül aktivitede dayanıklılık kaybı göstermiştir. FTIR spektroskopisi çalışmalarında tripsin jel ve agregatlarında 1637 cm-1 daki ?-yaprağı bantının yok olduğu görülmüştür. 1651 cm-1 daki ?-heliks bantının güçlenmesi tripsin jel ve agregatlarındaki alfa helix artişına karşılık gelmektedir. Iodoasetamidin jelleşmeyi geciktirmesi jelleşmede moleküller arası disülfit bantlarının önemine işaret etmektedir. Sonuç: Tripsin jelleşmesinin protein yapısındaki denatüreasyondan kaynaklandığı görülmüştür. Jel oluşumu protein ikincil yapısında ?-yaprağı kaybına paralel olarak ?-heliks oluşumu ve moleküller arası disülfit bağlarının oluşumu ile gerçekleşmektedir

Anahtar Kelime:

Konular: Biyokimya ve Moleküler Biyoloji

Tripsin çözelti, jel ve agregat formlarının organik çözgen içeren ve pH-tektiklenmiş faz geçişlerinde yapısal ve fonksiyonel incelenmesi

Öz:

In this study the effect of three different physicochemical parameters on pHtriggered gelation and aggregation of bovine pancreatic trypsin changes and structural and functional changes in these changes in alcohol-water mixtures were studied.Methods: Trypsin gelation times were studied using inverted tube method. Trypsin stability was studied using trypsin enzyme assay. Protein secondary structural changes were monitored using FTIR spectroscopy. Gel and aggregate macrostructures and morphologies were viewed using Scanning Electron Microscopy.Results: The solution phase was observed in the absence of both NaOH and CaCl2. The gel phase was observed in the absence of the either. The aggregate phase was observed in the presence of the both agents all depending on trypsin concentrations used. Trypsin stability studies showed that there were a nearly 53 and 32% specific activity losses after the gelation and aggregation processes. According to FTIR studies ?-sheet structure in 1637 cm-1 band disappeared in trypsin gel and trypsin aggregates. Increases in ?-helix structure in 1651 cm-1 in trypsin gel and aggregates were observed. Iodoacetamide delayed the gelation and prevented the aggregation indicating the importance of intermolecular disulfides in the both processes.Conclusion: Trypsin gelation was caused by the denaturation of the protein three dimensional structures. The gel and aggregate formation indicates a secondary structural change towards ?-helix structure formation at the expense of ?-sheet structure and formation of intermolecular disulfide bonds

Anahtar Kelime:

Konular: Biyokimya ve Moleküler Biyoloji

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	KARACİCEK B, Ceylan Ç (2015). Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. , 81 - 87.
Chicago	KARACİCEK Bilge,Ceylan Çağatay Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. (2015): 81 - 87.
MLA	KARACİCEK Bilge,Ceylan Çağatay Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. , 2015, ss.81 - 87.
AMA	KARACİCEK B,Ceylan Ç Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. . 2015; 81 - 87.
Vancouver	KARACİCEK B,Ceylan Ç Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. . 2015; 81 - 87.
IEEE	KARACİCEK B,Ceylan Ç "Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes." , ss.81 - 87, 2015.
ISNAD	KARACİCEK, Bilge - Ceylan, Çağatay. "Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes". (2015), 81-87.

APA	KARACİCEK B, Ceylan Ç (2015). Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. Türk Biyokimya Dergisi, 40(1), 81 - 87.
Chicago	KARACİCEK Bilge,Ceylan Çağatay Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. Türk Biyokimya Dergisi 40, no.1 (2015): 81 - 87.
MLA	KARACİCEK Bilge,Ceylan Çağatay Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. Türk Biyokimya Dergisi, vol.40, no.1, 2015, ss.81 - 87.
AMA	KARACİCEK B,Ceylan Ç Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. Türk Biyokimya Dergisi. 2015; 40(1): 81 - 87.
Vancouver	KARACİCEK B,Ceylan Ç Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes. Türk Biyokimya Dergisi. 2015; 40(1): 81 - 87.
IEEE	KARACİCEK B,Ceylan Ç "Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes." Türk Biyokimya Dergisi, 40, ss.81 - 87, 2015.
ISNAD	KARACİCEK, Bilge - Ceylan, Çağatay. "Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes". Türk Biyokimya Dergisi 40/1 (2015), 81-87.