• BMB Reports
• /
• 제42권7호
• /
• pp.418-420
• /
• 2009

We describe a method for producing polyclonal antibodies against peptide antigen cytochrome P450 1A2 and 3A4 using a tandem repeat of the epitope region and incorporation of proline residue between the repeated sequences. An ELISA assay revealed more efficient generation of polyclonal antibodies to tandem repeat peptide antigens than mono-epitope peptides. The incorporation of proline residues further stimulated antibody production.

• International Journal of Advanced Culture Technology
• /
• 제11권2호
• /
• pp.323-329
• /
• 2023

In this study, Proline pentamer model was used to investigate change in the dihedral angle, intramolecular hydrogen bonding and formation energies during structural optimization. L-Proline (LP, as an imino acid residue) pentamers having four conformation types [β: φ/ψ=t−/t+, α: φ/ψ=g−/g−, PP_II: φ/ψ=g−/t+ and Plike: φ/ψ= g−/g+] were carried out by QCC [B3LYP/6-31G(d,p)]. The optimized structure and formation energy were examined for designated structure. In LP, P-like and PP_II types did not change by optimization, and β types were transformed into PP_II having no H-bond independently of the designated ψ values. PP_II was more stable than P-like by about 2.2 kcal/mol/mu. The hydrogen bond distances of d2(4-6) type H-bonds were 1.94 - 2.00Å. In order to understand the processes of the transformations, the changes of φ/ψ, distances of NH-OC (d_NH/CO) and formation energies (ΔE, kcal/mol/mu) were examined.

• 한국식품영양과학회지
• /
• 제28권2호
• /
• pp.277-284
• /
• 1999

To confirm the food quality of conventionally processed fish extracts, protein quality of boiled crucian carp(Carassius carassius) and bastard halibut(Paralichthys olivaceus) extracts and their residues were evaluated. For the both fish extracts, some of the essential amino acids were lowered significantly but two times more proline and glycine were detected in extracts than those in raw fish meats. Boiling(100oC, 5 hours) caused 1.8(crucian carp)~2.4(bastard halibut) times more total free amino acid contents in fish extracts as compared to those in original fish meats. Taurine, glutamic acid, proline, lysine, and ammonia were the predominant free amino acids released in fish extracts. In vitro digestibility of boiled fish extracts were lower at a level of 4~6% than those of raw fish meats. Fish extraction residue had a higher in vitro digestibility and had a 60% lower level of TI than that of original fish meats. 18(bastard halibut)~ 24%(crucian carp) of available lysine was reduced in boiled fish extracts but a remarkable variation was not noted between extracts and residues. PERs and NPRs of fish extracts were significantly lower than those of casein, while those values of extraction residue were slightly higher as compared to those of control(ANRC casein). In vivo apparent digestibility exhibited a similar trend to in vitro digestibility. Hematological properties in serum of rat fed with fish extracts and residue were not changed significantly but the serum cholesterol concentration were reduced in rats fed fish extraction residue comparing with those of control. These results suggest that body weight loss due to fish extracts may not affect physiological changes.

• 한국식품과학회지
• /
• 제42권2호
• /
• pp.198-203
• /
• 2010

보리 맥아에는 2종의 $\alpha$-amylase isozyme(AMY1, AMY2)이 존재하며, 이들 효소는 80% 이상의 높은 아미노산 서열 상동성을 보이지만, calcium 의존성 등 효소의 작용특성은 서로 매우 다르다. 따라서 본 연구에서는 AMY2의 활성부위 중 2번째 $\beta\rightarrow\alpha$ loop에 존재하는 42번째 alanine 잔기를 saturation mutagenesis를 이용하여 다양한 아미노산으로 치환하고, 전분 분해활성이 증가한 돌연변이를 선발하였다. 결과적으로 alanine이 proline으로 치환된 AMY2-A42P의 경우에서만 발현도가 2배 증가하는 것을 확인하였으며, 특히 정제 과정에서의 회수율 또한 4배 증가하므로 향후 효소의 생산 및 활용에 유리할 것으로 판단하였다. 이 돌연변이 효소의 calcium 의존성 및 pH 안정성 등은 AMY2와 유사한 것으로 나타났으나, 각종 전분에 대한 기질특이성은 AMY1과 AMY2의 중간적인 특성으로 변화되었다. 결국 42번째 아미노산 잔기의 proline 치환에 의해 상대적으로 발현율이 높고 기질특이성이 변화된 AMY2 유사효소의 생산이 가능하였으며, 향후 이를 이용하여 분자진화기술 등 최신 효소공학적 방법론을 적용한 다양한 연구가 가능할 것으로 기대한다.

• 대한화학회지
• /
• 제22권6호
• /
• pp.386-395
• /
• 1978

여러 농도와 온도에서 Poly(trans-5-methyl-L-proline)을 trifluoroethanol 용매에서 정방향 변광회전, trifluoroethanol-n-butanol (1 : 4) 용매에서 역방향 변광회전을 시키면서 변광회전속도를 측정하였다. 이 두 방향의 변광회전 현상은 폴리머농도에 대하여 1차 반응이었다. 활성화에너지를 측정하기 위하여 변형된 Arrhenius식을 유도하였는데 이식은 물리적 성질과 농도와의 관계나 반응차수가 불확실한 반응에 이용할 수 있다. 정방향과 역방향 변광회전의 활성화에너지는 잔기몰당 각기 32.5와 33.5kcal이었고 이값은 polyproline 변광회전의 활성화에너지 (아미드결합의 공명에너지)보다 잔기몰당 10kcal가 높다. 이 과량의 활성화에너지는 폴리머아미드 결합이 시스-트란스 이성질화현상이 일어날 때 카르보닐기와 메틸기 사이에 생기는 입체장애에 의한 것이다.

• 한국미생물·생명공학회지
• /
• 제33권4호
• /
• pp.319-321
• /
• 2005

본 연구진은 phage display peptide library로부터 $TiO_{2}$ nanoparticle에 binding 하는 peptide를 선별하여 보고한바 있다. 이 중의 하나인 PEP9을 선택하여 alanine scanning mutagenesis를 통하여 mutant peptide를 display하는 phage를 제작하여 $TiO_{2}$에의 binding을 조사하였다. 그 결과, 4번 위치의 proline이 alanine으로 치환된 peptide의 경우 binding activity가 $10\%$로 감소하였고, 2번 valine, 3번 serine, 5번 isoleucine의 치환 peptide는 binding이 $40\%$로 감소하였다. 이러한 사실로 미루어볼 때, PEP9과 $TiO_{2}$ nanoparticle의 결합에는 2, 3, 4, 5번의 아미노산이 만들어내는 3차원적 구조가 중요한 역할을 하는 것으로 결론 내릴 수 있었다.

• BMB Reports
• /
• 제32권5호
• /
• pp.468-473
• /
• 1999

Nuclear Factor 1 (NF1) proteins are a family of transcriptional factors consisting of four different types: NF1-A, -B, -C, and -X. Some NF1 transcription factors contain a heptasequence motif, SPTSPSY, which is found as a repeat sequence in the carboxy terminal domain (CTD) of the largest subunit of RNA polymerase II. A similar heptasequence, SPTSPTY, is contained in rat liver NF1-A at a position between residues 469 and 475. In order to investigate the roles of the individual amino acids of the heptasequence of rat liver NF1-A in transcriptional activation, we systematically substituted single and multiple amino acid residues with alanine residue(s) and evaluated the transcriptional activities of the mutated NF1-A. Substitution of a single amino acid reduced transcriptional activity by 10 to 30%, except for the proline residue at position 473, whose substitution with alanine did not affect transcriptional activity. However, changes of all four serine and threonine residues to alanine or of the tyrosine residue along with the serine residue at position 469 to alanine reduced the activity to almost background levels. Our results indicate that multiple serine and threonine residues, rather than a single residue, may be involved in the modulation of the transcriptional activities of the factor. Involvement of the tyrosine residue is also implicated.

• Animal cells and systems
• /
• 제1권4호
• /
• pp.647-651
• /
• 1997

We have used a transient expression assay employing Drosophila S2 cells to study the transcriptional repression activity of a 27 amino acid residue-long repression domain FS1 which was generated by a frame-shift in a pair-rule gene, even-skipped of Drosophila melanogaster. In an attempt to define a minimal requirement for the repression activity, we constructed a series of truncation mutant forms of the FS1, fused to a heterologous GAL4 DNA-binding domain, and measured their activities. All of the mutant forms, including the GAL4-FS1 (5-23) which retains the smallest number (19) of amino acid residues of FS1, were found to repress an initiator, a minimal TATA-lacking promoter, in a GAL4-binding-site-dependent manner. These findings suggest that a 19 amino acid residue-long region, rich in proline and leucine residues, is a transcriptional repression domain and may interact with the general transcription machinery.

• BMB Reports
• /
• 제35권2호
• /
• pp.239-243
• /
• 2002

Angiotensin I that converts the enzyme (ACE) inhibitory peptide, Gly-Pro-Leu, previously purified and identified from the Alaskan pollack skin gelatin hydrolysate, were synthesized. In addition, the peptides Gly-Leu-Pro, Leu-Gly-Pro, Leu-Pro-Gly, Pro-Gly-Leu, Pro-Leu-Gly, Gly-Pro, and Pro-Leu, which consisted of glycine, proline, and leucine, were synthesized by the solid-phase method. The $IC_{50}$ values of each tripeptide - namely Leu-Gly-Pro, Gly-Leu-Pro, Gly-Pro-Leu, Pro-Leu-Gly, Leu-Pro-Gly, and Pro-Gly-Leu - were 0.72, 1.62, 2.65, 4.74, 5.73, and $13.93{\mu}M$, respectively. The ACE inhibitory activity of these tripeptides was higher than that of dipeptides, such as Gly-Pro and Pro-Leu with $IC_{50}$ values of 252.6 and $337.3\;{\mu}M$, respectively. Among the tripeptides, Leu-Gly-Pro and Gly-Leu-Pro had higher inhibitory activity than Gly-Pro-Leu that was isolated from the Alaskan pollack skin gelatin hydrolysate. Among the different types of tripeptides that were examined, the highest ACE inhibitory activity was observed for Leu-Gly-Pro. It had the leucine residue at the N-terminal and proline residue at the C-terminal.

• 한국식품영양과학회지
• /
• 제30권3호
• /
• pp.482-487
• /
• 2001

To find the possibility in utilizing the fish meat processing by-products, protein nutritional quality and textural properties of crucian carp extraction residue (CCER, feeze dired) incorporated into cookies were investigated. Moisture, ash and protein contents in cookies were increased with the higher residue treatments, but lipid contents were similar within all levels (3%, 9% and 15%). Major constitutional amino acids were revealed as glutamic acid, proline, leucine and arginine, and the sum of those amino acids was about 50% of total amino acid contents. Cookies with residue (CCER) had higher (80.74~84.50%) in vitro protein digestibility than standard cookies (83.32%), while slightly lower trypsin indigestible substrate (TIS) contents were showed in CCER containing cookies than control. CCER treatments resulted the decreased protein nutritional quality in C-PER (computed protein efficiency ratio) value from 2.41 (standard) to 1.15 (cinnamon flavored. 9% CCER), and those C-PER of all cookies were lower than ANRC casein (2.50). Lipophilic browning was developed steadily till 60 days storage and a significant (p<0.05) changes of browning ws not noteed between 60 days and 90 days storage. Color of cookies, expressed as L, a and b value, was significantly (p<0.05) lightened with the increased CCER. Similar trends by treatments were noted for hardness. Cookies containing 9% CCER were similar to control regarding textual and sensory properties.