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Signal Transduction of C-Terminal Phosphorylation Regions for Equine Luteinizing Hormone/Chorionic Gonadotropin Receptor (eLH/CGR)

  • Byambaragchaa, Munkhzaya;Joo, Hyo-Eun;Kim, Sang-Gwon;Kim, Yean-Ji;Park, Gyeong-Eun;Min, Kwan-Sik
    • Development and Reproduction
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    • v.26 no.1
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    • pp.1-12
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    • 2022
  • This study aimed to investigate the signal transduction of phosphorylation sites at the carboxyl (C)-terminal region of equine luteinizing hormone/chorionic gonadotropin receptor (eLH/CGR). The eLH/CGR has a large extracellular domain of glycoprotein hormone receptors within the G protein-coupled receptors. We constructed a mutant (eLH/CGR-t656) of eLH/CGR, in which the C-terminal cytoplasmic tail was truncated at the Phe656 residue, through polymerase chain reaction. The eLH/CGR-t656 removed 14 potential phosphorylation sites in the intracellular C-terminal region. The plasmids were transfected into Chinese hamster ovary (CHO)-K1 and PathHunter Parental cells expressing β-arrestin, and agonist-induced cAMP responsiveness was analyzed. In CHO-K1 cells, those expressing eLH/CGR-t656 were lower than those expressing eLH/CGR wild-type (eLH/CGR-wt). The EC50 of the eLH/CGR-t656 mutant was approximately 72.2% of the expression observed in eLH/CGR-wt. The maximal response in eLH/CGR-t656 also decreased to approximately 43% of that observed in eLH/CGR-wt. However, in PathHunter Parental cells, cAMP activity and maximal response of the eLH/CGR-t656 mutant were approximately 173.5% and 100.8%, respectively, of that of eLH/CGR-wt. These results provide evidence that the signal transduction of C-terminal phosphorylation in eLH/CGR plays a pivotal role in CHO-K1 cells. The cAMP level was recovered in PathHunter Parental cells expressing β-arrestin. We suggest that the signal transduction of the C-terminal region phosphorylation sites is remarkably different depending on the cells expressing β-arrestin in CHO-K1 cells.

Satisfaction with Neighborhood Environments for Dog Walking and Exercise (반려견 산책 및 운동을 위한 근린환경 인식 및 만족도 연구)

  • Song, Jin-Gyeoung;Lee, Jeong-Yeon;Lee, Hyung-Sook
    • Journal of the Korean Institute of Landscape Architecture
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    • v.50 no.4
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    • pp.53-62
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    • 2022
  • Animal companionship is gaining popularity in Korea, and interest in dog walking is also increasing as a strategy to maintain the physical and mental health of dogs and dog owners. The purpose of this study was to investigate the perception and satisfaction of dog owners with the neighborhood environments for dog walking and to examine the willingness to pay (WTP) for an off-leash dog park. Of a total of 249 online survey respondents, 47.8% said they take a walk at least once a day and mainly walk around nearby neighborhoods or parks. Respondents showed a high attachment to their dogs and recognized that dog walks help increase physical activity and maintain the health of owners and their dogs. The satisfaction with neighborhood environments, such as attractive views and the connection of walking trails, was low, and there was a high demand for off-leash dog parks. The truncated mean WTP was estimated to be 85,618 Korean Won, and the result of regression analysis and t-test revealed that walking frequency and the satisfaction with dog walking facilities were statistically significant in the WTP. In order to encourage dog walking to promote physical activity and health, more attention needs to be devoted to improving neighborhood environments for dog walking.

Elementary preservice teachers' conceptions of a generating line and exploration of teaching methods (모선에 대한 초등 예비교사의 인식과 지도 방안 탐색)

  • Jinam Hwang
    • The Mathematical Education
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    • v.63 no.3
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    • pp.451-466
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    • 2024
  • This study investigated elementary preservice teachers' conceptions of a generating line, an ambiguous concept in school mathematics. The preservice teachers' conceptions of a generating line can be classified into four types: (a) only cones have generating lines, (b) only cones and cylinders have generating lines, (c) solids of revolution have generating lines, (d) straight lines on the lateral surface are generating lines. 22.1% of all preservice teachers believed that only cones have generating lines, and most of them followed the definition of a generating line presented in elementary mathematics textbooks. The conception that only cones and cylinders have generating lines was the least investigated. However, since there were instances where generating lines were defined with the use of a director curve, it became important to explore topics more thoroughly, such as generating lines of a truncated cone. 27.9% of all preservice teachers believed that solids of revolution have generating lines. This conception was marked by differing opinions on whether spheres also have generating lines. The conception that straight lines on the lateral surface are generating lines was the most frequently investigated. This conception differs from the traditional view in school mathematics because it suggests using a director curve to define generating lines. Based on these analysis results, the researcher developed specific teaching methods that considered both subject matter knowledge and pedagogical content knowledge for preservice teachers. In addition, the researcher proposed a consensus definition of a generating line in mathematics education.

Analysis of Hydrocarbon Trap in the Southwestern Margin of the Ulleung Basin, East Sea (동해 울릉분지 남서주변부의 탄화수소 트랩 분석)

  • Lee, Minwoo;Kang, Moo-Hee;Yoon, Youngho;Yi, Bo-Yeon;Kim, Kyong-O;Kim, Jinho;Park, Myong-ho;Lee, Keumsuk
    • Economic and Environmental Geology
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    • v.48 no.4
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    • pp.301-312
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    • 2015
  • A commercial gas field was found in the southwestern continental shelf of the Ulleung Basin, East Sea in the late 1990s. To develop additional gas field, an exploration well was drilled through the coarse infill of submarine canyon near the gas field, but it was uneconomic to develop hydrocarbons. Using newly acquired deep seismic reflection and previous well data, we have identified additional geological structure which has hydrocarbon potentials below submarine canyons in the southwestern margin of the basin. Based on the interpretation of the deep seismic reflection and well data, the sequences of the study area can be classified into the syn-rift megasequence(MS1), post-rift megasequence(MS2), syn-compressional megasequence(MS3), and post-compressional megasequence(MS4) in relation to the tectonic events. MS1, deposited simultaneously with the basin formation before the middle Miocene, is characterized by chaotic seismic facies with low- to moderate-amplitude and low frequency reflections. MS2 comprises laterally continuous, low- to moderate-amplitude reflections, showing progradational stacking patterns due to high rates of sediment supply during basin expansion in the middle Miocene. MS3 is mainly composed of continuous reflections with high amplitude and moderate- to high-frequency which are interpreted as coarse-grained sediments. The coarse-grained sediments of MS3 sequence is widely truncated by several submarine canyons which filled with fine-grained sediment of MS4 to form a stratigraphic trap of hydrocarbon. Therefore, the reservoir and seal of the hydrocarbon trap in the study area are coarse-grained sediment of MS3 and submarine canyon filled with fine-grained sediment of MS4, respectively. A flat-spot seismic anomaly, which may indicate the presence of hydrocarbon, is observed within the stratigraphic trap.

Expression of Peroxiredoxin and Thioredoxin in Human Lung Cancer and Paired Normal Lung (인체의 폐암과 정상 폐조직에서 Peroxiredoxin 및 Thioredoxin의 발현 양상)

  • Kim, Young Sun;Park, Joo Hun;Lee, Hye Lim;Shim, Jin Young;Choi, Young In;Oh, Yoon Jung;Shin, Seung Soo;Choi, Young Hwa;Park, Kwang Joo;Park, Rae Woong;Hwang, Sung Chul
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.2
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    • pp.142-150
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    • 2005
  • Background : Continuous growth stimulation by various factors, as well as chronic oxidative stress, may co-exist in many solid tumors, such as lung cancer. A new family of antioxidant proteins, the peroxiredoxins (Prxs), have been implicated in the regulation of many cellular processes, including cell proliferation, differentiation and apoptosis. However, a real pathophysiological significance of Prx proteins, especially in lung disease, has not been sufficiently defined. Therefore, this study was conducted to investigate the distribution and expression of various Prx isoforms in lung cancer and other pulmonary conditions. Method : Patients diagnosed with lung cancer, and who underwent surgery at the Ajou Medical Center, were enrolled. The expressions of Prxs, Thioredoxin (Trx) and Thioredoxin reductase (TR) were analyzed using proteomic techniques and the subcellular localization of Prx proteins was studied using immunohistochemistry on normal mouse lung tissue. Result : Immunohistochemical staining has shown the isoforms of Prx I, II, III and V are predominantly expressed in bronchial and alveolar lining epithelia, as well as in the alveolar macrophages of the normal mouse lung. The isoforms of Prx I and III, and thioredoxin were also found to be over-expressed in the lung cancer tissues compared to their paired normal lung controls. There was also an increased amount of the oxidized form of Prx I, as well as a putative truncated form of Prx III, in the lung cancer samples when analyzed using 2-dimensional electrophoresis. In addition, a 43 kDa intermediate molecular weight protein band, and other high molecular weight bands of over 20 kDa, recognized by the anti-Prx I antibody, were present in the tissue extracts of lung cancer patients on 1-Dimensional electrophoresis, which require further investigation. Conclusion : The over-expressions of Prx I and III, and Trx in human lung cancer tissue, as well as their possible chaperoning function, may represent an attempt by tumor cells to adjust to their microenvironment in a manner advantageous to their survival and proliferation, while maintaining their malignant potential.

Flowering and Morphological Responses of Petunia and Pansy as Influenced by Lamp Type and Lighting Period to Provide Long Days (장일처리 광원의 종류 및 광조사 시간에 따른 페튜니아와 팬지의 개화 및 형태학적 반응)

  • Oh, Wook;Runkle, Erik S.
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.207-219
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    • 2016
  • Incandescent (INC) lamps have been commonly used to promote flowering of long-day (LD) plants during short-day (SD) seasons, but production of INC lamps has been prohibited due to their low energy efficiency. One of the light sources replacing INC lamps is a compact fluorescent lamp (CFL). This study was carried out to compare the flowering and morphological responses of LD annuals grown in a controlled environment greenhouse at $20^{\circ}C$ with a truncated 9-h SD and a 2- or 4-h night interruption (NI) or 6-h day extension (DE) provided by lighting from INC lamps, CFLs, or a combination of the two (INC + CFLs), in which red (R) to far-red (FR) ratios were 0.60, 8.46, and 0.91, respectively, and their PPFDs were $2.3{\pm}0.3{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. After 12 weeks of treatment, $Petunia{\times}hybrida$ 'Wave Purple' plants did not flower under the SD photoperiod whereas 100% flowered under all of the LD treatments. Flowering was more rapid under the INC or INC + CFL lighting treatments compared to CFL and DE, and 4-h NI enhanced flowering compared to 2-h NI. In addition, plants under DE and 4h-NI generally flowered earlier than under 2-h NI. All petunia 'Single Dreams Red' plants flowered within 65 days after treatment, and flowering was hastened by some LD lighting regimens and lamp types. Plants under DE and 4h-NI generally flowered earlier than under 2-h NI INC or INC + CFL compared to FL, and flowering time under INC 6-h DE was earliest. In addition, INC lighting promoted stem elongation of both petunia cultivars. In both pansy (Viola${\times}wittrockiana$) 'Coiossus Yellow' and 'Delta Blue Blotch', LD treatments, especially using INC lamps, promoted flowering whereas the lighting period had little influence on days to flowering. Therefore, INC or INC + CFL with lower R:FR promoted flowering and stem extension and the promoting effect was larger with longer lighting periods. These results suggest that CFLs can be used to provide LDs to promote the flowering of petunia and pansy and to reduce stem elongation, although the promoting effect on flowering is sometimes less than that of INC lamps alone.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Oxidative Stress-dependent Structural and Functional Regulation of 2-cysteine Peroxiredoxins In Eukaryotes Including Plant Cells (산화 스트레스에 의존한 식물 및 진핵세포 2-시스테인 퍼록시레독신의 기능 조절)

  • Jang, Ho-Hee;Kim, Sun-Young;Lee, Sang-Yeol
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.1-9
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    • 2006
  • Peroxiredoxins (Prxs) are ubiquitously distributed and play important functions in diverse cellular signaling systems. The proteins are largely classified into three groups, such as typical 2-Cys Prx, atypical 2-Cys Prx, and 1-Cys Prx, that are distinguished by their catalytic mechanisms and number of Cys residues. From the three classes of Prxs, the typical 2-Cys Prx containing the two-conserved Cys residues at its N-terminus and C-terminus catalyzes $H_2O_2$ with the use of thioredoxin (Trx) as an electron donor. During the catalytic cycle, the N-terminal Cys residue undergoes a peroxide-dependent oxidation to sulfenic acid, which can be further oxidized to sulfinic acid at the presence of high concentrations of $H_2O_2$ and a Trx system containing Trx, Trx reductase, and NADPH. The sulfinic acid form of 2-Cys Prx is reduced by the action of sulfiredoxin which requires ATP as an energy source. Under the strong oxidative or heat shock stress conditions, 2-Cys Prx in eukaryotes rapidly switches its protein structure from low-molecular-weight species to high-molecular-weight protein structures. In accordance with its structural changes, the protein concomitantly triggers functional switching from a peroxidase to a molecular chaperone, which can protect its substrate denaturation from external stress. In addition to its N-terminal active site, the C-terminal domain including 'YF-motif' of 2-Cys Prx plays a critical role in the structural changes. Therefore, the C-terminal truncated 2-Cys Prxs are not able to regulate their protein structures and highly resistant to $H_2O_2$-dependent hyperoxidation, suggesting that the reaction is guided by the peroxidatic Cys residue. Based on the results, it may be concluded that the peroxidatic Cys of 2-Cys Prx acts as an '$H_2O_2$-sensor' in the cells. The oxidative stress-dependent regulation of 2-Cys Prx provides a means of defense systems in cells to adapt stress conditions by activating intracellular defense signaling pathways. Particularly, 2-Cys Prxs in plants are localized in chloroplasts with a dynamic protein structure. The protein undergoes conformational changes again oxidative stress. Depending on a redox-potential of the chloroplasts, the plant 2-Cys Prx forms super-molecular weight protein structures, which attach to the thylakoid membranes in a reversible manner.

Investigation on Inhibitory Effect of ErmSF N-Terminal End Region Peptide on ErmSF Methyltansferase Activity In Vivo Through Development of Co-Expression System of Two Different Proteins in One Cell (서로 다른 두 단백질의 세포 내 동시 발현 체계의 개발을 통한 ErmSF에서 특이적으로 발견되는 N-Terminal End Region (NTER)을 포함하는 펩타이드의 생체내에서의 ErmSF 활성 억제 효과 검색)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.200-208
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    • 2011
  • Most problematic antibiotic resistance mechanism for MLS (macrolide-lincosamide-streptogramn B) antibiotics encountered in clinical practice is mono- or dimethylation of specific adenine residue at 2058 (E. coli coordinate) of 23S rRNA which is performed by Erm (erythromycin ribosome resistance) protein through which bacterial ribosomes reduce the affinity to the antibiotics and become resistant to them. ErmSF is one of the four gene products produced by Streptomyces fradiae to be resistant to its own antibiotic, tylosin. Unlike other Erm proteins, ErmSF harbors idiosyncratic long N-terminal end region (NTER) 25% of which is comprised of arginine well known to interact with RNA. Furthermore, NTER was found to be important because when it was truncated, most of the enzyme activity was lost. Based on these facts, capability of NTER peptide to inhibit the enzymatic activity of ErmSF was sought. For this, expression system for two different proteins to be expressed in one cell was developed. In this system, two plasmids, pET23b and pACYC184 have unique replication origins to be compatible with each other in a cell. And expression system harboring promoter, ribosome binding site and transcription termination signal is identical but disparate amount of protein could be expressed according to the copy number of each vector, 15 for pACYC and 40 for pET23b. Expression of NTER peptide in pET23b together with ErmSF in pACYC 184 in E. coli successfully gave more amounts of NTER than ErmSF but no inhibitory effects were observed suggesting that there should be dynamicity in interaction between ErmSF and rRNA rather than simple and fixed binding to each other in methylation of 23S rRNA by ErmSF.

A Variable Latency Goldschmidt's Floating Point Number Square Root Computation (가변 시간 골드스미트 부동소수점 제곱근 계산기)

  • Kim, Sung-Gi;Song, Hong-Bok;Cho, Gyeong-Yeon
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.9 no.1
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    • pp.188-198
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    • 2005
  • The Goldschmidt iterative algorithm for finding a floating point square root calculated it by performing a fixed number of multiplications. In this paper, a variable latency Goldschmidt's square root algorithm is proposed, that performs multiplications a variable number of times until the error becomes smaller than a given value. To find the square root of a floating point number F, the algorithm repeats the following operations: $R_i=\frac{3-e_r-X_i}{2},\;X_{i+1}=X_i{\times}R^2_i,\;Y_{i+1}=Y_i{\times}R_i,\;i{\in}\{{0,1,2,{\ldots},n-1} }}'$with the initial value is $'\;X_0=Y_0=T^2{\times}F,\;T=\frac{1}{\sqrt {F}}+e_t\;'$. The bits to the right of p fractional bits in intermediate multiplication results are truncated, and this truncation error is less than $'e_r=2^{-p}'$. The value of p is 28 for the single precision floating point, and 58 for the doubel precision floating point. Let $'X_i=1{\pm}e_i'$, there is $'\;X_{i+1}=1-e_{i+1},\;where\;'\;e_{i+1}<\frac{3e^2_i}{4}{\mp}\frac{e^3_i}{4}+4e_{r}'$. If '|X_i-1|<2^{\frac{-p+2}{2}}\;'$ is true, $'\;e_{i+1}<8e_r\;'$ is less than the smallest number which is representable by floating point number. So, $\sqrt{F}$ is approximate to $'\;\frac{Y_{i+1}}{T}\;'$. Since the number of multiplications performed by the proposed algorithm is dependent on the input values, the average number of multiplications per an operation is derived from many reciprocal square root tables ($T=\frac{1}{\sqrt{F}}+e_i$) with varying sizes. The superiority of this algorithm is proved by comparing this average number with the fixed number of multiplications of the conventional algorithm. Since the proposed algorithm only performs the multiplications until the error gets smaller than a given value, it can be used to improve the performance of a square root unit. Also, it can be used to construct optimized approximate reciprocal square root tables. The results of this paper can be applied to many areas that utilize floating point numbers, such as digital signal processing, computer graphics, multimedia, scientific computing, etc.